Taq Polymerase
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Enzyme activities: T4 DNA Ligase catalyzes the formation of phosphodiester bonds between neighbouring 3'-hydroxyl and 5'-phosphate ends in double-stranded DNA. Sticky- and blunt-ended DNA fragments are ligated. Single-stranded nicks in double-stranded DNA are also closed.
Appropriate ligation buffer, 10x concentrated: Tris/HCl, 660 mmol/L; MgCl2, 50 mmol/L; DTT, 50 mmol/L; ATP, 10 mmol/ L, pH 7.5 at +20°C (Note: ATP is not stable).
pH optimum: 7.2-7.8 Divalent ion requirement: Mg2+ Inactivation: After 10 minutes heat denaturation at +65°C ligase activity is stopped
實驗流程
Enzyme activities: T4 DNA Ligase catalyzes the formation of phosphodiester bonds between neighbouring 3'-hydroxyl and 5'-phosphate ends in double-stranded DNA. Sticky- and blunt-ended DNA fragments are ligated. Single-stranded nicks in double-stranded DNA are also closed.
Appropriate ligation buffer, 10x concentrated: Tris/HCl, 660 mmol/L; MgCl2, 50 mmol/L; DTT, 50 mmol/L; ATP, 10 mmol/ L, pH 7.5 at +20°C (Note: ATP is not stable).
pH optimum: 7.2-7.8 Divalent ion requirement: Mg2+ Inactivation: After 10 minutes heat denaturation at +65°C ligase activity is stopped
實驗數據
Enzyme activities: T4 DNA Ligase catalyzes the formation of phosphodiester bonds between neighbouring 3'-hydroxyl and 5'-phosphate ends in double-stranded DNA. Sticky- and blunt-ended DNA fragments are ligated. Single-stranded nicks in double-stranded DNA are also closed.
Appropriate ligation buffer, 10x concentrated: Tris/HCl, 660 mmol/L; MgCl2, 50 mmol/L; DTT, 50 mmol/L; ATP, 10 mmol/ L, pH 7.5 at +20°C (Note: ATP is not stable).
pH optimum: 7.2-7.8 Divalent ion requirement: Mg2+ Inactivation: After 10 minutes heat denaturation at +65°C ligase activity is stopped
產品原理
Enzyme activities: T4 DNA Ligase catalyzes the formation of phosphodiester bonds between neighbouring 3'-hydroxyl and 5'-phosphate ends in double-stranded DNA. Sticky- and blunt-ended DNA fragments are ligated. Single-stranded nicks in double-stranded DNA are also closed.
Appropriate ligation buffer, 10x concentrated: Tris/HCl, 660 mmol/L; MgCl2, 50 mmol/L; DTT, 50 mmol/L; ATP, 10 mmol/ L, pH 7.5 at +20°C (Note: ATP is not stable).
pH optimum: 7.2-7.8 Divalent ion requirement: Mg2+ Inactivation: After 10 minutes heat denaturation at +65°C ligase activity is stopped