商品编号


EG-1053



品牌


Excellgen



公司


Excellgen, Inc.



公司分类


Ligases




20,000 Units

商品信息



Description

T4 RNA Ligase 2 truncated catalyzes phosphodiester bond formation between a pre-adenylated 5′ phosphate (DNA or RNA) and the 3′ hydroxyl of RNA. The truncated enzyme containing the first 249 amino acids is unable to adenylate the 5? end of the substrate, thus it can not ligate the phosphorylated 5? end of RNA or DNA to the 3? end of RNA. Such a property is useful for certain applications such as linker ligations with pre-adenylated 5′ DNA to 3′ hydroxyl RNA, such as microRNAs. This truncated enzyme significantly reduces unwanted background ligation as it can only use adenylated primers.



Applications



Ligate a pre-adenylated DNA or RNA sequence tag to any RNA 3?-end

Join a single stranded adenylated primer to small RNAs for
CDN
A library creation

Join a single stranded adenylated primer to RNA for strand-specific
CDN
A library construction





Source

Purified from a recombinant
E. coli
strain carrying the T4 phage RNA Ligase 2 Rnl2 (gp24.1) (1-249) gene



Components



T4 RNA Ligase 2 truncated
: 200,000 units/ml in storage buffer: 10 mM Tris-HCl, 100 mM NaCl, 0.1 mM DTT, 0.1 mM EDTA, 50% glycerol, pH 7.5 @ 25°C

10 X T4 RNA Ligase 2 truncated Buffer
: 500 mM Tris-HCl, 100 mM MgCl
2
, 50 mM DTT, pH 7.6 @ 25°C





Unit Definition

One unit is defined as the amount of enzyme required to ligate 50% of 0.4 ?g of an equimolar mix of a single-stranded 5′ FAM-labeled 17-mer RNA to the 5′ pre-adenylated end of a 18-mer DNA when both 17-mers are annealed to a complementary 35-mer DNA strand in 20 μL 1X reaction buffer following a 30 minute incubation at 37°C.



Quality Control



The absence of endo-, exodeoxyribonucleases, phosphatases, and ribonucleases confirmed by appropriate quality tests.

Functionally tested for the capacity to join cohesive- and blunt-end DNA fragments.





Storage

-20°C.