M0208 FAQs for Taq DNA Ligase, Ligases, Intl, NEB
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Taq DNA Ligase FAQ

Q1: Is Taq DNA Ligase used for a special technique?
Q2: How much ligation occurs at mismatches when using Taq DNA Ligase?
Q3: How many temperature cycles will Taq DNA Ligase survive?
Q4: Can Taq DNA Ligase be used for cloning?
Q5: What is the molecular weight of Taq DNA Ligase?
Q6: What is the specific activity of Taq DNA Ligase?
Q7: Why is the Taq DNA Ligase buffer brown?
Q8: Does Taq DNA Ligase require NAD?
Q9: What is the activity of Taq DNA Ligase in other NEBuffers?
Q10: What is the activity of Taq DNA Ligase at various temperatures?
Q11: What is the stability of Taq DNA Ligase at 95C?
Q12: What is the stability of Taq DNA Ligase at room temperature?

Q1: Is Taq DNA Ligase used for a special technique?

A1: Yes. It is used in LCR (ligase chain reaction) and LDR (ligase detection reaction). In general a single base change can be detected by two probes that meet with the terminal base of one probe pairing with the base being analyzed. If the terminal base pairs with the template, ligase can join the two probes. If the terminal base does not pair, ligase will not join the probes. Since the ligase is thermostable the reaction can be cycled and the ligation products amplified.


Q2: How much ligation occurs at mismatches when using Taq DNA Ligase?

A2: The maximum seen is 1.3 % ligation at mismatches.


Q3: How many temperature cycles will Taq DNA Ligase survive?

A3: At least 30 cycles.


Q4: Can Taq DNA Ligase be used for cloning?

A4: Taq DNA Ligase will not ligate all types of ends.We recommend using T4 DNA Ligase (
NEB# M0202) for cloning.


Q5: What is the molecular weight of Taq DNA Ligase?

A5: The adenylated form is 81kD and the nonadenylated form is 78 kD.


Q6: What is the specific activity of Taq DNA Ligase?

A6: The specific activity is 1.67 x 106 units/mg and the protein concentration is 0.3 pmol/μl.


Q7: Why is the Taq DNA Ligase buffer brown?

A7: The DTT and NAD react and turn brown if the buffer is stored at less than -70C. The brown buffer will still work.


Q8: Does Taq DNA Ligase require NAD?

A8: Yes. There is enough NAD bound to the enzyme so it will work to some extent in buffers that do not contain NAD.


Q9: What is the activity of Taq DNA Ligase in other NEBuffers?

A9: NEBuffer 2: 10,000 units/ml
NEBuffer 3: 8-10,000 units/ml
NEBuffer 4: 12-16,000 units/ml
Taq DNA Ligase: 40,000 units/ml
TaqI restriction enzyme unique: 16,000 units/ml


Q10: What is the activity of Taq DNA Ligase at various temperatures?

A10: 4C: 2,000 units/ml
16C: 8-10,000 units/ml
25C: 12,000 units/ml
37C: 20,000 units/ml
45C: 40,000 units/ml
55C: 40,000 units/ml
65C: 64,000 units/ml
75C: 10,000 units/ml
85C: 3-4,000 units/ml
95C: < 500 units/ml


Q11: What is the stability of Taq DNA Ligase at 95C?

A11: 0 min: 64,000 units/ml
1 min: 40-50,000 units/ml
5 min: 40-50,000 units/ml
10 min: 32,000 units/ml
15 min: 32,000 units/ml
20 min: 32,000 units/ml
30 min: 32,000 units/ml
45 min: 16,000 units/ml
60 min: 16,000 units/ml


Q12: What is the stability of Taq DNA Ligase at room temperature?

A12: Over one week.