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T4 DNA/重组T4 DNA 连接酶
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T4 DNA/重组T4 DNA 连接酶
产品编号：ENZ-286
产品规格：20,000IU/100,000IU/500,000IU
Synonyms DNA ligase 4, EC 6.5.1.1, DNA ligase IV, Polydeoxyribonucleotide synthase [ATP] 4. Description T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5? -phosphate ａnd 3? -hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end ａnd cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids. Source Escherichia Colilambda lysogen NM 989. Physical Appearance Sterile filtered liquid formulation having a concentration of 167,000 U/ml. Formulation 50mM Tris-HCl （pH 7.8 at 25°C）, 10mM MgCl2, 10mM DTT, 1mM ATP, 25 μg/ml BSA ａnd DNA （0.1 to 1 μm in 5′ termini）. ?ptimal ligation occurs at 16oC. Storage Buffer 50mM KCl, 10mM Tris-HCl （pH 7.4）, 0.1mM EDTA, 1mM DTT, 200 μg/ml BSA ａnd 50% glycerol. Store at -20oC. Stability Two years when stored at –20°C, 2 weeks at 4°C. Unit Defenition 1. One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of ? DNA （5′ DNA termini concentration of 0.12 μM, 300- μg/ml） in a total reaction volume of 20 ?l in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer.
2.One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmol of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37°C. Biological Activity One Weiss unit is equivalent to circa 67 cohesive-end ligation units.
T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration is > 200mM.
·Ligation of blunt-ended ａnd single-base pair overhang fragments requires about 50 times as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt-end ligation may be enhanced by addition of PEG 4000 （10% w/v final concentration） or hexamine chloride, or by reducing the ATP concentration to 50?M.
·To dilute T4 DNA Ligase that will subsequently be stored at –200C, 50% glycerol storage buffer should be used; to dilute for immediate use, 1x T4 DNA Ligase reaction buffer can be used. Heat Inactivation T4 DNA Ligase can be inactivated by incubation at 65oC for 10 minutes. Quality Control Purified free of contaminating endonucleases ａnd exonucleases. Each lot of T4 DNA ligase is also tested in a mock cloning assay, which reveals any damage to the ligated DNA termini. Greater than 99.9% of the termini remain undamaged in this assay. Exonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of a mixture of single ａnd double-stranded [3H] E. coli DNA （200,000 cpm/?g） for 4 hours at 37°C released < 0.3% of the total radioactivity. Endonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of ?X174 RF I DNA for 4 hours at 37°C resulted in < 5% conversion to RFII as determined by agarose gel electrophoresis. Nuclease Activity Incubation of 13,000 units for 18 hours in assay buffer （without ATP） with Hind III fragments of ? DNA yielded a clear ａnd sharp banding pattern on agarose gels. Applications Cloning of restriction fragments.
joining linkers ａnd adapters to blunt-ended DNA.
T4 DNA/重组T4 DNA 连接酶
产品编号：ENZ-286
产品规格：20,000IU/100,000IU/500,000IU
Synonyms DNA ligase 4, EC 6.5.1.1, DNA ligase IV, Polydeoxyribonucleotide synthase [ATP] 4. Description T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5? -phosphate ａnd 3? -hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end ａnd cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids. Source Escherichia Colilambda lysogen NM 989. Physical Appearance Sterile filtered liquid formulation having a concentration of 167,000 U/ml. Formulation 50mM Tris-HCl （pH 7.8 at 25°C）, 10mM MgCl2, 10mM DTT, 1mM ATP, 25 μg/ml BSA ａnd DNA （0.1 to 1 μm in 5′ termini）. ?ptimal ligation occurs at 16oC. Storage Buffer 50mM KCl, 10mM Tris-HCl （pH 7.4）, 0.1mM EDTA, 1mM DTT, 200 μg/ml BSA ａnd 50% glycerol. Store at -20oC. Stability Two years when stored at –20°C, 2 weeks at 4°C. Unit Defenition 1. One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of ? DNA （5′ DNA termini concentration of 0.12 μM, 300- μg/ml） in a total reaction volume of 20 ?l in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer.
2.One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmol of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37°C. Biological Activity One Weiss unit is equivalent to circa 67 cohesive-end ligation units.
T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration is > 200mM.
·Ligation of blunt-ended ａnd single-base pair overhang fragments requires about 50 times as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt-end ligation may be enhanced by addition of PEG 4000 （10% w/v final concentration） or hexamine chloride, or by reducing the ATP concentration to 50?M.
·To dilute T4 DNA Ligase that will subsequently be stored at –200C, 50% glycerol storage buffer should be used; to dilute for immediate use, 1x T4 DNA Ligase reaction buffer can be used. Heat Inactivation T4 DNA Ligase can be inactivated by incubation at 65oC for 10 minutes. Quality Control Purified free of contaminating endonucleases ａnd exonucleases. Each lot of T4 DNA ligase is also tested in a mock cloning assay, which reveals any damage to the ligated DNA termini. Greater than 99.9% of the termini remain undamaged in this assay. Exonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of a mixture of single ａnd double-stranded [3H] E. coli DNA （200,000 cpm/?g） for 4 hours at 37°C released < 0.3% of the total radioactivity. Endonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of ?X174 RF I DNA for 4 hours at 37°C resulted in < 5% conversion to RFII as determined by agarose gel electrophoresis. Nuclease Activity Incubation of 13,000 units for 18 hours in assay buffer （without ATP） with Hind III fragments of ? DNA yielded a clear ａnd sharp banding pattern on agarose gels. Applications Cloning of restriction fragments.
joining linkers ａnd adapters to blunt-ended DNA.
T4 DNA/重组T4 DNA 连接酶
产品编号：ENZ-286
产品规格：20,000IU/100,000IU/500,000IU
Synonyms DNA ligase 4, EC 6.5.1.1, DNA ligase IV, Polydeoxyribonucleotide synthase [ATP] 4. Description T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5? -phosphate ａnd 3? -hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end ａnd cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids. Source Escherichia Colilambda lysogen NM 989. Physical Appearance Sterile filtered liquid formulation having a concentration of 167,000 U/ml. Formulation 50mM Tris-HCl （pH 7.8 at 25°C）, 10mM MgCl2, 10mM DTT, 1mM ATP, 25 μg/ml BSA ａnd DNA （0.1 to 1 μm in 5′ termini）. ?ptimal ligation occurs at 16oC. Storage Buffer 50mM KCl, 10mM Tris-HCl （pH 7.4）, 0.1mM EDTA, 1mM DTT, 200 μg/ml BSA ａnd 50% glycerol. Store at -20oC. Stability Two years when stored at –20°C, 2 weeks at 4°C. Unit Defenition 1. One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of ? DNA （5′ DNA termini concentration of 0.12 μM, 300- μg/ml） in a total reaction volume of 20 ?l in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer.
2.One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmol of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37°C. Biological Activity One Weiss unit is equivalent to circa 67 cohesive-end ligation units.
T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration is > 200mM.
·Ligation of blunt-ended ａnd single-base pair overhang fragments requires about 50 times as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt-end ligation may be enhanced by addition of PEG 4000 （10% w/v final concentration） or hexamine chloride, or by reducing the ATP concentration to 50?M.
·To dilute T4 DNA Ligase that will subsequently be stored at –200C, 50% glycerol storage buffer should be used; to dilute for immediate use, 1x T4 DNA Ligase reaction buffer can be used. Heat Inactivation T4 DNA Ligase can be inactivated by incubation at 65oC for 10 minutes. Quality Control Purified free of contaminating endonucleases ａnd exonucleases. Each lot of T4 DNA ligase is also tested in a mock cloning assay, which reveals any damage to the ligated DNA termini. Greater than 99.9% of the termini remain undamaged in this assay. Exonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of a mixture of single ａnd double-stranded [3H] E. coli DNA （200,000 cpm/?g） for 4 hours at 37°C released < 0.3% of the total radioactivity. Endonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of ?X174 RF I DNA for 4 hours at 37°C resulted in < 5% conversion to RFII as determined by agarose gel electrophoresis. Nuclease Activity Incubation of 13,000 units for 18 hours in assay buffer （without ATP） with Hind III fragments of ? DNA yielded a clear ａnd sharp banding pattern on agarose gels. Applications Cloning of restriction fragments.
joining linkers ａnd adapters to blunt-ended DNA.
T4 DNA/重组T4 DNA 连接酶
产品编号：ENZ-286
产品规格：20,000IU/100,000IU/500,000IU
Synonyms DNA ligase 4, EC 6.5.1.1, DNA ligase IV, Polydeoxyribonucleotide synthase [ATP] 4. Description T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5? -phosphate ａnd 3? -hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end ａnd cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids. Source Escherichia Colilambda lysogen NM 989. Physical Appearance Sterile filtered liquid formulation having a concentration of 167,000 U/ml. Formulation 50mM Tris-HCl （pH 7.8 at 25°C）, 10mM MgCl2, 10mM DTT, 1mM ATP, 25 μg/ml BSA ａnd DNA （0.1 to 1 μm in 5′ termini）. ?ptimal ligation occurs at 16oC. Storage Buffer 50mM KCl, 10mM Tris-HCl （pH 7.4）, 0.1mM EDTA, 1mM DTT, 200 μg/ml BSA ａnd 50% glycerol. Store at -20oC. Stability Two years when stored at –20°C, 2 weeks at 4°C. Unit Defenition 1. One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of ? DNA （5′ DNA termini concentration of 0.12 μM, 300- μg/ml） in a total reaction volume of 20 ?l in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer.
2.One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmol of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37°C. Biological Activity One Weiss unit is equivalent to circa 67 cohesive-end ligation units.
T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration is > 200mM.
·Ligation of blunt-ended ａnd single-base pair overhang fragments requires about 50 times as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt-end ligation may be enhanced by addition of PEG 4000 （10% w/v final concentration） or hexamine chloride, or by reducing the ATP concentration to 50?M.
·To dilute T4 DNA Ligase that will subsequently be stored at –200C, 50% glycerol storage buffer should be used; to dilute for immediate use, 1x T4 DNA Ligase reaction buffer can be used. Heat Inactivation T4 DNA Ligase can be inactivated by incubation at 65oC for 10 minutes. Quality Control Purified free of contaminating endonucleases ａnd exonucleases. Each lot of T4 DNA ligase is also tested in a mock cloning assay, which reveals any damage to the ligated DNA termini. Greater than 99.9% of the termini remain undamaged in this assay. Exonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of a mixture of single ａnd double-stranded [3H] E. coli DNA （200,000 cpm/?g） for 4 hours at 37°C released < 0.3% of the total radioactivity. Endonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of ?X174 RF I DNA for 4 hours at 37°C resulted in < 5% conversion to RFII as determined by agarose gel electrophoresis. Nuclease Activity Incubation of 13,000 units for 18 hours in assay buffer （without ATP） with Hind III fragments of ? DNA yielded a clear ａnd sharp banding pattern on agarose gels. Applications Cloning of restriction fragments.
joining linkers ａnd adapters to blunt-ended DNA.
T4 DNA/重组T4 DNA 连接酶
产品编号：ENZ-286
产品规格：20,000IU/100,000IU/500,000IU
Synonyms DNA ligase 4, EC 6.5.1.1, DNA ligase IV, Polydeoxyribonucleotide synthase [ATP] 4. Description T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5? -phosphate ａnd 3? -hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end ａnd cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids. Source Escherichia Colilambda lysogen NM 989. Physical Appearance Sterile filtered liquid formulation having a concentration of 167,000 U/ml. Formulation 50mM Tris-HCl （pH 7.8 at 25°C）, 10mM MgCl2, 10mM DTT, 1mM ATP, 25 μg/ml BSA ａnd DNA （0.1 to 1 μm in 5′ termini）. ?ptimal ligation occurs at 16oC. Storage Buffer 50mM KCl, 10mM Tris-HCl （pH 7.4）, 0.1mM EDTA, 1mM DTT, 200 μg/ml BSA ａnd 50% glycerol. Store at -20oC. Stability Two years when stored at –20°C, 2 weeks at 4°C. Unit Defenition 1. One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of ? DNA （5′ DNA termini concentration of 0.12 μM, 300- μg/ml） in a total reaction volume of 20 ?l in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer.
2.One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmol of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37°C. Biological Activity One Weiss unit is equivalent to circa 67 cohesive-end ligation units.
T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration is > 200mM.
·Ligation of blunt-ended ａnd single-base pair overhang fragments requires about 50 times as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt-end ligation may be enhanced by addition of PEG 4000 （10% w/v final concentration） or hexamine chloride, or by reducing the ATP concentration to 50?M.
·To dilute T4 DNA Ligase that will subsequently be stored at –200C, 50% glycerol storage buffer should be used; to dilute for immediate use, 1x T4 DNA Ligase reaction buffer can be used. Heat Inactivation T4 DNA Ligase can be inactivated by incubation at 65oC for 10 minutes. Quality Control Purified free of contaminating endonucleases ａnd exonucleases. Each lot of T4 DNA ligase is also tested in a mock cloning assay, which reveals any damage to the ligated DNA termini. Greater than 99.9% of the termini remain undamaged in this assay. Exonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of a mixture of single ａnd double-stranded [3H] E. coli DNA （200,000 cpm/?g） for 4 hours at 37°C released < 0.3% of the total radioactivity. Endonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of ?X174 RF I DNA for 4 hours at 37°C resulted in < 5% conversion to RFII as determined by agarose gel electrophoresis. Nuclease Activity Incubation of 13,000 units for 18 hours in assay buffer （without ATP） with Hind III fragments of ? DNA yielded a clear ａnd sharp banding pattern on agarose gels. Applications Cloning of restriction fragments.
joining linkers ａnd adapters to blunt-ended DNA.
T4 DNA/重组T4 DNA 连接酶
产品编号：ENZ-286
产品规格：20,000IU/100,000IU/500,000IU
Synonyms DNA ligase 4, EC 6.5.1.1, DNA ligase IV, Polydeoxyribonucleotide synthase [ATP] 4. Description T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5? -phosphate ａnd 3? -hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end ａnd cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids. Source Escherichia Colilambda lysogen NM 989. Physical Appearance Sterile filtered liquid formulation having a concentration of 167,000 U/ml. Formulation 50mM Tris-HCl （pH 7.8 at 25°C）, 10mM MgCl2, 10mM DTT, 1mM ATP, 25 μg/ml BSA ａnd DNA （0.1 to 1 μm in 5′ termini）. ?ptimal ligation occurs at 16oC. Storage Buffer 50mM KCl, 10mM Tris-HCl （pH 7.4）, 0.1mM EDTA, 1mM DTT, 200 μg/ml BSA ａnd 50% glycerol. Store at -20oC. Stability Two years when stored at –20°C, 2 weeks at 4°C. Unit Defenition 1. One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of ? DNA （5′ DNA termini concentration of 0.12 μM, 300- μg/ml） in a total reaction volume of 20 ?l in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer.
2.One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmol of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37°C. Biological Activity One Weiss unit is equivalent to circa 67 cohesive-end ligation units.
T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration is > 200mM.
·Ligation of blunt-ended ａnd single-base pair overhang fragments requires about 50 times as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt-end ligation may be enhanced by addition of PEG 4000 （10% w/v final concentration） or hexamine chloride, or by reducing the ATP concentration to 50?M.
·To dilute T4 DNA Ligase that will subsequently be stored at –200C, 50% glycerol storage buffer should be used; to dilute for immediate use, 1x T4 DNA Ligase reaction buffer can be used. Heat Inactivation T4 DNA Ligase can be inactivated by incubation at 65oC for 10 minutes. Quality Control Purified free of contaminating endonucleases ａnd exonucleases. Each lot of T4 DNA ligase is also tested in a mock cloning assay, which reveals any damage to the ligated DNA termini. Greater than 99.9% of the termini remain undamaged in this assay. Exonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of a mixture of single ａnd double-stranded [3H] E. coli DNA （200,000 cpm/?g） for 4 hours at 37°C released < 0.3% of the total radioactivity. Endonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of ?X174 RF I DNA for 4 hours at 37°C resulted in < 5% conversion to RFII as determined by agarose gel electrophoresis. Nuclease Activity Incubation of 13,000 units for 18 hours in assay buffer （without ATP） with Hind III fragments of ? DNA yielded a clear ａnd sharp banding pattern on agarose gels. Applications Cloning of restriction fragments.
joining linkers ａnd adapters to blunt-ended DNA.
T4 DNA/重组T4 DNA 连接酶
产品编号：ENZ-286
产品规格：20,000IU/100,000IU/500,000IU
Synonyms DNA ligase 4, EC 6.5.1.1, DNA ligase IV, Polydeoxyribonucleotide synthase [ATP] 4. Description T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5? -phosphate ａnd 3? -hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end ａnd cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids. Source Escherichia Colilambda lysogen NM 989. Physical Appearance Sterile filtered liquid formulation having a concentration of 167,000 U/ml. Formulation 50mM Tris-HCl （pH 7.8 at 25°C）, 10mM MgCl2, 10mM DTT, 1mM ATP, 25 μg/ml BSA ａnd DNA （0.1 to 1 μm in 5′ termini）. ?ptimal ligation occurs at 16oC. Storage Buffer 50mM KCl, 10mM Tris-HCl （pH 7.4）, 0.1mM EDTA, 1mM DTT, 200 μg/ml BSA ａnd 50% glycerol. Store at -20oC. Stability Two years when stored at –20°C, 2 weeks at 4°C. Unit Defenition 1. One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of ? DNA （5′ DNA termini concentration of 0.12 μM, 300- μg/ml） in a total reaction volume of 20 ?l in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer.
2.One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmol of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37°C. Biological Activity One Weiss unit is equivalent to circa 67 cohesive-end ligation units.
T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration is > 200mM.
·Ligation of blunt-ended ａnd single-base pair overhang fragments requires about 50 times as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt-end ligation may be enhanced by addition of PEG 4000 （10% w/v final concentration） or hexamine chloride, or by reducing the ATP concentration to 50?M.
·To dilute T4 DNA Ligase that will subsequently be stored at –200C, 50% glycerol storage buffer should be used; to dilute for immediate use, 1x T4 DNA Ligase reaction buffer can be used. Heat Inactivation T4 DNA Ligase can be inactivated by incubation at 65oC for 10 minutes. Quality Control Purified free of contaminating endonucleases ａnd exonucleases. Each lot of T4 DNA ligase is also tested in a mock cloning assay, which reveals any damage to the ligated DNA termini. Greater than 99.9% of the termini remain undamaged in this assay. Exonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of a mixture of single ａnd double-stranded [3H] E. coli DNA （200,000 cpm/?g） for 4 hours at 37°C released < 0.3% of the total radioactivity. Endonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of ?X174 RF I DNA for 4 hours at 37°C resulted in < 5% conversion to RFII as determined by agarose gel electrophoresis. Nuclease Activity Incubation of 13,000 units for 18 hours in assay buffer （without ATP） with Hind III fragments of ? DNA yielded a clear ａnd sharp banding pattern on agarose gels. Applications Cloning of restriction fragments.
joining linkers ａnd adapters to blunt-ended DNA.
T4 DNA/重组T4 DNA 连接酶
产品编号：ENZ-286
产品规格：20,000IU/100,000IU/500,000IU
Synonyms DNA ligase 4, EC 6.5.1.1, DNA ligase IV, Polydeoxyribonucleotide synthase [ATP] 4. Description T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5? -phosphate ａnd 3? -hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end ａnd cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids. Source Escherichia Colilambda lysogen NM 989. Physical Appearance Sterile filtered liquid formulation having a concentration of 167,000 U/ml. Formulation 50mM Tris-HCl （pH 7.8 at 25°C）, 10mM MgCl2, 10mM DTT, 1mM ATP, 25 μg/ml BSA ａnd DNA （0.1 to 1 μm in 5′ termini）. ?ptimal ligation occurs at 16oC. Storage Buffer 50mM KCl, 10mM Tris-HCl （pH 7.4）, 0.1mM EDTA, 1mM DTT, 200 μg/ml BSA ａnd 50% glycerol. Store at -20oC. Stability Two years when stored at –20°C, 2 weeks at 4°C. Unit Defenition 1. One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of ? DNA （5′ DNA termini concentration of 0.12 μM, 300- μg/ml） in a total reaction volume of 20 ?l in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer.
2.One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmol of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37°C. Biological Activity One Weiss unit is equivalent to circa 67 cohesive-end ligation units.
T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration is > 200mM.
·Ligation of blunt-ended ａnd single-base pair overhang fragments requires about 50 times as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt-end ligation may be enhanced by addition of PEG 4000 （10% w/v final concentration） or hexamine chloride, or by reducing the ATP concentration to 50?M.
·To dilute T4 DNA Ligase that will subsequently be stored at –200C, 50% glycerol storage buffer should be used; to dilute for immediate use, 1x T4 DNA Ligase reaction buffer can be used. Heat Inactivation T4 DNA Ligase can be inactivated by incubation at 65oC for 10 minutes. Quality Control Purified free of contaminating endonucleases ａnd exonucleases. Each lot of T4 DNA ligase is also tested in a mock cloning assay, which reveals any damage to the ligated DNA termini. Greater than 99.9% of the termini remain undamaged in this assay. Exonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of a mixture of single ａnd double-stranded [3H] E. coli DNA （200,000 cpm/?g） for 4 hours at 37°C released < 0.3% of the total radioactivity. Endonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of ?X174 RF I DNA for 4 hours at 37°C resulted in < 5% conversion to RFII as determined by agarose gel electrophoresis. Nuclease Activity Incubation of 13,000 units for 18 hours in assay buffer （without ATP） with Hind III fragments of ? DNA yielded a clear ａnd sharp banding pattern on agarose gels. Applications Cloning of restriction fragments.
joining linkers ａnd adapters to blunt-ended DNA.
T4 DNA/重组T4 DNA 连接酶
产品编号：ENZ-286
产品规格：20,000IU/100,000IU/500,000IU
Synonyms DNA ligase 4, EC 6.5.1.1, DNA ligase IV, Polydeoxyribonucleotide synthase [ATP] 4. Description T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5? -phosphate ａnd 3? -hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end ａnd cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids. Source Escherichia Colilambda lysogen NM 989. Physical Appearance Sterile filtered liquid formulation having a concentration of 167,000 U/ml. Formulation 50mM Tris-HCl （pH 7.8 at 25°C）, 10mM MgCl2, 10mM DTT, 1mM ATP, 25 μg/ml BSA ａnd DNA （0.1 to 1 μm in 5′ termini）. ?ptimal ligation occurs at 16oC. Storage Buffer 50mM KCl, 10mM Tris-HCl （pH 7.4）, 0.1mM EDTA, 1mM DTT, 200 μg/ml BSA ａnd 50% glycerol. Store at -20oC. Stability Two years when stored at –20°C, 2 weeks at 4°C. Unit Defenition 1. One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of ? DNA （5′ DNA termini concentration of 0.12 μM, 300- μg/ml） in a total reaction volume of 20 ?l in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer.
2.One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmol of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37°C. Biological Activity One Weiss unit is equivalent to circa 67 cohesive-end ligation units.
T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration is > 200mM.
·Ligation of blunt-ended ａnd single-base pair overhang fragments requires about 50 times as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt-end ligation may be enhanced by addition of PEG 4000 （10% w/v final concentration） or hexamine chloride, or by reducing the ATP concentration to 50?M.
·To dilute T4 DNA Ligase that will subsequently be stored at –200C, 50% glycerol storage buffer should be used; to dilute for immediate use, 1x T4 DNA Ligase reaction buffer can be used. Heat Inactivation T4 DNA Ligase can be inactivated by incubation at 65oC for 10 minutes. Quality Control Purified free of contaminating endonucleases ａnd exonucleases. Each lot of T4 DNA ligase is also tested in a mock cloning assay, which reveals any damage to the ligated DNA termini. Greater than 99.9% of the termini remain undamaged in this assay. Exonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of a mixture of single ａnd double-stranded [3H] E. coli DNA （200,000 cpm/?g） for 4 hours at 37°C released < 0.3% of the total radioactivity. Endonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of ?X174 RF I DNA for 4 hours at 37°C resulted in < 5% conversion to RFII as determined by agarose gel electrophoresis. Nuclease Activity Incubation of 13,000 units for 18 hours in assay buffer （without ATP） with Hind III fragments of ? DNA yielded a clear ａnd sharp banding pattern on agarose gels. Applications Cloning of restriction fragments.
joining linkers ａnd adapters to blunt-ended DNA.
T4 DNA/重组T4 DNA 连接酶
产品编号：ENZ-286
产品规格：20,000IU/100,000IU/500,000IU
Synonyms DNA ligase 4, EC 6.5.1.1, DNA ligase IV, Polydeoxyribonucleotide synthase [ATP] 4. Description T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5? -phosphate ａnd 3? -hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end ａnd cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids. Source Escherichia Colilambda lysogen NM 989. Physical Appearance Sterile filtered liquid formulation having a concentration of 167,000 U/ml. Formulation 50mM Tris-HCl （pH 7.8 at 25°C）, 10mM MgCl2, 10mM DTT, 1mM ATP, 25 μg/ml BSA ａnd DNA （0.1 to 1 μm in 5′ termini）. ?ptimal ligation occurs at 16oC. Storage Buffer 50mM KCl, 10mM Tris-HCl （pH 7.4）, 0.1mM EDTA, 1mM DTT, 200 μg/ml BSA ａnd 50% glycerol. Store at -20oC. Stability Two years when stored at –20°C, 2 weeks at 4°C. Unit Defenition 1. One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of ? DNA （5′ DNA termini concentration of 0.12 μM, 300- μg/ml） in a total reaction volume of 20 ?l in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer.
2.One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmol of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37°C. Biological Activity One Weiss unit is equivalent to circa 67 cohesive-end ligation units.
T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration is > 200mM.
·Ligation of blunt-ended ａnd single-base pair overhang fragments requires about 50 times as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt-end ligation may be enhanced by addition of PEG 4000 （10% w/v final concentration） or hexamine chloride, or by reducing the ATP concentration to 50?M.
·To dilute T4 DNA Ligase that will subsequently be stored at –200C, 50% glycerol storage buffer should be used; to dilute for immediate use, 1x T4 DNA Ligase reaction buffer can be used. Heat Inactivation T4 DNA Ligase can be inactivated by incubation at 65oC for 10 minutes. Quality Control Purified free of contaminating endonucleases ａnd exonucleases. Each lot of T4 DNA ligase is also tested in a mock cloning assay, which reveals any damage to the ligated DNA termini. Greater than 99.9% of the termini remain undamaged in this assay. Exonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of a mixture of single ａnd double-stranded [3H] E. coli DNA （200,000 cpm/?g） for 4 hours at 37°C released < 0.3% of the total radioactivity. Endonuclease Activity Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of ?X174 RF I DNA for 4 hours at 37°C resulted in < 5% conversion to RFII as determined by agarose gel electrophoresis. Nuclease Activity Incubation of 13,000 units for 18 hours in assay buffer （without ATP） with Hind III fragments of ? DNA yielded a clear ａnd sharp banding pattern on agarose gels. Applications Cloning of restriction fragments.
joining linkers ａnd adapters to blunt-ended DNA.
产品规格：20,000IU/100,000IU/500,000IU
| Synonyms | DNA ligase 4, EC 6.5.1.1, DNA ligase IV, Polydeoxyribonucleotide synthase [ATP] 4. |
| Description | T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5? -phosphate ａnd 3? -hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end ａnd cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids. |
| Source | Escherichia Colilambda lysogen NM 989. |
| Physical Appearance | Sterile filtered liquid formulation having a concentration of 167,000 U/ml. |
| Formulation | 50mM Tris-HCl （pH 7.8 at 25°C）, 10mM MgCl2, 10mM DTT, 1mM ATP, 25 μg/ml BSA ａnd DNA （0.1 to 1 μm in 5′ termini）. ?ptimal ligation occurs at 16oC. |
| Storage Buffer | 50mM KCl, 10mM Tris-HCl （pH 7.4）, 0.1mM EDTA, 1mM DTT, 200 μg/ml BSA ａnd 50% glycerol. Store at -20oC. |
| Stability | Two years when stored at –20°C, 2 weeks at 4°C. |
| Unit Defenition | 1. One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of ? DNA （5′ DNA termini concentration of 0.12 μM, 300- μg/ml） in a total reaction volume of 20 ?l in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer. 2.One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmol of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37°C. |
| Biological Activity | One Weiss unit is equivalent to circa 67 cohesive-end ligation units. T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration is > 200mM. ·Ligation of blunt-ended ａnd single-base pair overhang fragments requires about 50 times as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt-end ligation may be enhanced by addition of PEG 4000 （10% w/v final concentration） or hexamine chloride, or by reducing the ATP concentration to 50?M. ·To dilute T4 DNA Ligase that will subsequently be stored at –200C, 50% glycerol storage buffer should be used; to dilute for immediate use, 1x T4 DNA Ligase reaction buffer can be used. |
| Heat Inactivation | T4 DNA Ligase can be inactivated by incubation at 65oC for 10 minutes. |
| Quality Control | Purified free of contaminating endonucleases ａnd exonucleases. Each lot of T4 DNA ligase is also tested in a mock cloning assay, which reveals any damage to the ligated DNA termini. Greater than 99.9% of the termini remain undamaged in this assay. |
| Exonuclease Activity | Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of a mixture of single ａnd double-stranded [3H] E. coli DNA （200,000 cpm/?g） for 4 hours at 37°C released < 0.3% of the total radioactivity. |
| Endonuclease Activity | Incubation of a 50?l reaction containing 13,000 units of T4 DNA Ligase with 1?g of ?X174 RF I DNA for 4 hours at 37°C resulted in < 5% conversion to RFII as determined by agarose gel electrophoresis. |
| Nuclease Activity | Incubation of 13,000 units for 18 hours in assay buffer （without ATP） with Hind III fragments of ? DNA yielded a clear ａnd sharp banding pattern on agarose gels. |
| Applications | Cloning of restriction fragments. joining linkers ａnd adapters to blunt-ended DNA. |
T4,DNA,重组T4,DNA,连接酶
