T4 RNA Ligase 2, truncated K227Q
T4 RNA Ligase 2, truncated K227Q ligates the pre-adenylated 5´ end of DNA or RNA to the 3´ end of RNA.
- Reduced lysyl adenylation activity
- Isolated from a recombinant source
- Supplied with 10X Reaction Buffer
- Requires 5' pre-adenylated RNA or DNA for ligation
- Tested for the absence of endonucleases, exonucleases, RNases and phosphatases
- Not sure which ligase to choose? Refer to our DNA and RNA Ligase Properties Chart
T4 RNA Ligase 2, truncated K227Q (T4 Rnl2tr K227Q) specifically ligates the pre-adenylated 5´ end of DNA or RNA to the 3´ OH end of RNA. The enzyme does not use ATP for ligation but requires pre-adenylated linkers.
T4 Rnl2tr K227Q is a point mutant of T4 RNA Ligase 2, truncated (NEB# M0242) . Mutation of K227 in T4 RNA Ligase 2 reduces enzyme lysyl adenylation (1). This mutation further reduces the formation of undesired ligation products (concatemers and circles) by T4 Rnl2tr (2), possibly by reducing the trace activity of T4 Rnl2tr in transfer of adenylyl groups from linkers to the 5´-phosphates of input RNAs.
The exclusion of ATP, use of pre-adenylated linkers, and the reduced enzyme lysyl adenylation activity provide the lowest possible background in ligation reactions. This enzyme has been used for optimized linker ligation for the cloning of microRNAs (2).
Product SourceT4 Rnl2tr K227Q is expressed as an MBP fusion from a plasmid in E. coli which encodes the first 249 amino acids of the full length T4 RNA Ligase 2 with a lysine to glutamine mutation at position 227.
The following reagents are supplied with this product:
Store at (°C) Concentration T4 RNA Ligase Reaction Buffer -20 10 X PEG 8000 -20 50 %
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