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商品編號


M0319S-10 reactions



品牌


紐英倫(NEB)



公司


New England Biolabs



公司分類(lèi)


DNA Modifying Enzymes




10 reactions


Concentration

20 μM

商品信息

Description:
Thermo
stable 5? App DNA/RNALigase is a point mutant of catalytic lysine of RNAligase from
Methanobacterium
Thermo
autotrophicum
(1). This enzyme is ATP independent. Itrequires a 5? pre-adenylated linker for ligation tothe 3?-OH end of either RNA or single strandedDNA (ssDNA). The enzyme is also active in ligationof RNA with 2?-O-methylated 3? end to 5?-adenylatedlinkers (1). The optimal temperature forligation reaction is 60–65°C (2). The mutant ligaseis unable to adenylate the 5?-phosphate of RNA orssDNA, which reduces the formation of undesiredligation products (concatemers and circles).
The
ABI
lity of the ligase to function at 65°C mightreduce the constraints of RNA secondary structurein RNA ligation experiments.
Product Source
Thermo
stable 5? App DNA/RNA Ligase isexpressed as His-tag fusion in
E. coli
.
Reagents Supplied
The following reagents are supplied with this product:
Store at (°C)
Concentration
NEB
uffer? 1
-20
10X
MnCl
2
10X
Notes:
For optimal ligation of ssDNA toadenylated DNA linkers, we recommend using
NEB
uffer 1 supplemented with manganese. Forligation of ssRNA to adenylated DNA linkers, justuse
NEB
uffer 1. Heat inactivation or Proteinase Ktreatment is required to release RNA bound by theligase.