商品编号


EG-1052



品牌


Excellgen



公司


Excellgen, Inc.



公司分类


Ligases




250 Units

商品信息



Description

T4 RNA Ligase 2, also known as T4 Rnl2 (gp24.1), has both intermolecular and intramolecular RNA strand joining activity. T4 RNA Ligase 2 is much more active joining nicks on double stranded RNA than on joining the ends of single stranded RNA. Ligation of single-stranded RNA substrates generates either intramolecular or intermolecular products. The enzyme requires an adjacent 5? phosphate and 3? OH for ligation. The enzyme can also ligate the 3? OH of RNA to the 5? phosphate of DNA in a double stranded structure. The non-ligated strand of the duplex can be either RNA or DNA. T4 RNA ligase 2 requires ATP for activity unless the substrate is preadenylated on the 5′ end.



Applications



Ligate a nick in dsRNA

Ligate the 3? OH of RNA to the 5? phosphate of DNA in a double stranded structure





Source


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




Components



T4 RNA Ligase 2
: 30,000 U/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 Buffer
: 500 mM Tris-HCl, 100 mM MgCl
2
, 50 mM DTT, 10 mM ATP, 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′ phosphorylated end of a 18-mer DNA when both strands are annealed to a complementary 35-mer DNA strand in 20 ?L at 37°C for 30 minutes.




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.