Modifying Enzymes
Sigma Aldrich Fine Chemicals Biosciences Hyaluronidase from Bovine Testes | 37326-33-3 | MFCD00131351 | 100mg
Hyaluronidase from Bovine Testes | 37326-33-3 | MFCD00131351 | 100mg
Sigma Aldrich Fine Chemicals Biosciences Hyaluronidase from Bovine Testes | 37326-33-3 | MFCD00131351 | 100mg
Hyaluronidase from Bovine Testes | 37326-33-3 | MFCD00131351 | 100mg
New England Biolabs, Inc. Thermostable 5' App DNA/RNA Ligase – 10 reactions
Thermostable 5' App DNA/RNA Ligase is a point mutant of catalytic lysine of RNA ligase from Methanobacterium thermoautotrophicum. This enzyme is ATP independent. It requires a 5' pre-adenylated linker for ligation to the 3' -OH end of either RNA or single stranded DNA (ssDNA). The enzyme is also active in ligation of RNA with 2' -O-methylated 3 end to 5-adenylated linkers. The optimal temperature for ligation reaction is 60-65°C. The mutant ligase is unable to adenylate the 5' -phosphate of RNA or ssDNA, which reduces the formation of undesired ligation products (concatemers and circles). The ability of the ligase to function at 65°C might reduce the constraints of RNA secondary structure in RNA ligation experiments.
New England Biolabs, Inc. SplintR Ligase – 1250 units
SplintR Ligase efficiently catalyzes the ligation of adjacent, single-stranded DNA splinted by a complementary RNA strand. This activity may enable novel approaches for characterization of miRNAs and mRNAs, including SNPs. The robust activity of the enzyme and its affinity for RNA-splinted DNA substrates (apparent Km = 1 nM) enable sub-nanomolar detection of unique RNA species within a complex mixture, making SplintR ligase a superior choice for demanding RNA detection technologies. The activity is enhanced at higher temperatures (up to 37°C), and by supplementation with 5 mM Mn2+. The reaction is inhibited by salt concentrations in excess of 100 mM. The enzyme tolerates all base pair combinations at the ligation junction, but is partially inhibited by dC/G and dG/C base pairs at the donor (phosphorylated) side ligation junction, particularly when the +2 base was also a C/G base pair.
New England Biolabs, Inc. T4 RNA Ligase 2, truncated KQ – 2000 units
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 . Mutation of K227 in T4 RNA Ligase 2 reduces enzyme lysyl adenylation. K227Q reduces the formation of undesired ligation products (concatemers and circles) by T4 Rnl2tr, by reducing the trace activity of T4 Rnl2tr in transfer of adenylyl groups from linkers to the 5-phosphates of input RNAs. Mutation of R55K in T4 Rnl2tr K227Q increases the ligation activity of the enzyme to levels similar to T4 Rnl2tr. 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 .
Zymo Research Corporation dsDNA Shearase™ Plus 50 U (1 U/ul)
dsDNA Shearase Plus is an endonuclease that cleaves phosphodiester bonds in DNA to yield oligonucleotides with 5’-phosphate and 3’-hydroxyl termini. It has a particularly strong preference for dsDNA and generates random-ended DNA fragments of the desired size in a single step. This enzyme is compatible with low volume inputs, thus minimizing sample loss.
