Chinese Name	连接酶
Price	32/60U
Storage	-20 ℃
ALIAS	T4 DNA connection kit

 

 

Features:

High -efficiency connecting viscosity end Heping End

t/a cloning

DSDNA cutting repairs

Construct a high -abundance cloning library [

123]

Connection of RNA and DNA

Overview:

This enzyme catalyzes dual chain DNA or RNA adjacent 5′-phosphate end and 3′-hydroxymal end to form phosphate two. Essence. This enzyme can not only catalyze the connection between the flat or sticky end of the end, but also repair the single -chain cutting in the dual -chain DNA, RNA or DNA/RNA hybridization dual chain.

Source:

Purified from E. Coli C600 PCL857 PPLC28 LIG8.

Reaction condition:

1x T4 DNA connecting enzyme reaction buffer

[50 mm tris-hCl (pH 7.5 @ 25 ° C), 10 mm mgCl2, 10 mm DTTTT , 1 mm atp]. The end concentration of the DNA 5 ′ end is 0.1-1.0 μm, and it is cultivated at 16 ° C.

Heat loss: 65 ° C 10 minutes.

The unit definition (viscosity end activity unit):

1 unit refers to the 20 μL 1x T4 DNA connection enzyme reaction buffer, under 16 ° C reaction conditions, 30 minutes can make 50% of 50% The λDNA fragment [5 ′ end concentration of Hind Ⅲ is 0.12 μm (300 μg/ml)] to connect the enzyme amount required.

Note:

Different from the E. Coli DNA connecting enzymes, the auxiliary factor of the T4 DNA connecting enzyme is ATP.

The diluted T4 DNA connection enzyme should be kept in -20 ° C and 50% glycerin storage buffer (diluted buffer A, NEB #B8001). If you use it immediately after dilution, you can also dilute with 1x T4 DNA connection enzyme reaction buffer.

As long as 1 mm ATP is added in the reaction system, the connection reaction can be connected to the four restricted internal cutase buffer provided in NEB or

T4 DNA polyetroinucleotide kinase kinase kinase kinase kinase Perform in the reaction buffer.

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