T4 RNA Ligase 2 (truncated) (RNL2)

Description

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. By not having an extra ATP in the reaction, the amount of ligation between random RNA molecules is dramatically reduced. Unlike the full length T4 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 your plan to sequence miRNAs or to perform directional mRNA-Seq, the truncated ligase 2 will help to enhance your library preparations.
Figure, Bioanalyzer trace of ligation product with T4 RNA Ligase 2 (dsRNA Ligase). The enzyme can efficiently ligate 3′ OH of RNA fragment to the 5′ phosphate of DNA fragment in a double stranded structure. Red color trace: New lot enzyme reaction; Blue color trace: Previous lot enzyme reaction (shelf life was near 12 months).

Features:
– Developed for Next-Generation RNA Sequencing applications
– Efficiency of ligation at nearly 100%
– Increase ability to identify miRNAs

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

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 DTT
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.

Specific Activity: 1,200 U/μg

Recommended Storage Condition: -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.

Additional information

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