T4 RNA Ligase 2 (truncated) (RNL2)

Description    
MCLAB’s truncated  T4 RNA Ligase 2 was developed specifically for demanding Next-Generation RNA Sequencing applications.  The truncated  ligase 2 specifically ligates the adenylated 5´ end of an adapter to the 3´ end of RNA. The enzyme does not require ATP for ligation but does need an adenylated substrate, which dramatically reduces the amount of ligation between random RNA molecules.  Unlike the full lengthT4 RNA  ligase 2, the truncated ligase does not ligate the phosphorylated 5´ end of RNA or DNA without the adenylated substrate making it an excellent choice for small RNA library preparation. Whether you plan to sequence miRNAs or perform directional mRNA-Seq,  The truncated ligase 2 will enhance your library preparations.

Features
- develped for  Next-Generation RNA Sequencing applications
- Efficiency of ligation nears 100%
- Increase ability to identify miRNAs

Source:
Purified from an E. coli strain carrying T4 RNA Ligase 2 truncated (1-249) overproducing plasmid.

Specific Activity:
20,000U/mg

Heat Inactivation:
65°C for 20 minutes

Reaction Conditions:
1X T4 RNA Ligase Reaction Buffer
Incubate at 25°C.

1X T4 RNA Ligase Reaction Buffer:
50 mM Tris-HCl
10 mM MgCl2
1 mM Dithiothreitol
pH 7.5 @ 25°C

Storage Conditions:
10 mM Tris-HCl (pH 7.5)
100 mM NaCl
0.1 mM EDTA
 0.1 mM DTT and 50% Glycerol

Unit Definition:
One unit is defined as the amount of the enzyme required to give 50% ligation of a 17-mer adenylated oligonucleotide to a purified control RNA template in 30 minutes at 37°C.

 Recommended Storage Conditions:
-20°C

Quality Control:
Enzyme preparations are routinely assessed for relative purity, activity and absence of RNase and DNase

References:

1. Jayaprakash, A. D., Jabado O., Brown, B. D. and Sachidanandam, R. (Sept 2, 2011),  Nuc Acid Res, 1–12.
2. Ho, C.K. et al. (2004) Structure, 12, 327-339.
3. Ho, C.K. and Shuman, S. (2002) Proc. Natl.Acad.Sci. USA, 99, 12709-12714.