T4 RNA Ligase 2, truncated KQ
T4 RNA Ligase 2, truncated KQ 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
Avoiding RNase Contamination
T4 RNA Ligase 2, truncated KQ (T4 Rnl2tr R55K, K227Q) specifically ligates the preadenylated 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 R55K, K227Q is a double-point mutant of T4 RNA Ligase 2, truncated (NEB #M0242). Mutation of K227 in T4 RNA Ligase 2 reduces enzyme lysyl adenylation (1). K227Q reduces the formation of undesired ligation products (concatemers and circles) by T4 Rnl2tr (2,3), by reducing the trace activity of T4 Rnl2tr in transfer of adenylyl groups from linkers to the 5´-phosphates of input RNAs (3). Mutation of R55K in T4 Rnl2tr K227Q increases the ligation activity of the enzyme to levels similar to T4 Rnl2tr (3).
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 high-throughput sequencing library construction of small RNA (4).
Product SourceT4 Rnl2tr R55K, 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 an arginine to lysine and a lysine to glutamine mutations at positions 55 and 227, respectively.
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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