AIR™ Ligase

T4 RNA Ligase 2 (RNL2)

Catalog# Product Name Quantity US List Price
NOVA-512105 AIR™ Ligase 5,000 Units
$144 Buy Now

 

 

  • Truncation makes downstream assays more robust
  • Efficiency of ligation nears 100%
  • Increase ability to identify miRNAs 

 

Bioo Scientific’s AIR™ Ligase was developed specifically for demanding Next-Generation RNA Sequencing applications. As can be seen in the figure below, AIR Ligase 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. AIR Ligase is a truncated version of T4 RNA Ligase 2. Unlike the full length ligase, AIR 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 microRNAs or perform directional mRNA-Seq, AIR Ligase will enhance your library preparations. 


Select Publications that Cite Use of AIR Ligase:

Campo, S., Gilbert, K. B. and Carrington, J. C. (2016) Small RNA-Based Antiviral Defense in the Phytopathogenic Fungus Colletotrichum higginsianum. PLOS Pathogens. doi: 10.1371/journal.ppat.1005640.

Haenni, S. et al. (March 2012) Analysis of C. elegans intestinal gene expression and polyadenylation by fluorescence-activated nuclei sorting and 3'-end-seq. Nuc Acid Res, 40:6

Hoque, M., Ji, Z. et al. (2012) Analysis of alternative cleavage and polyadenylation by 3′ region extraction and deep sequencing. Nature Methods. DOI:10.1038/NMETHMETHMETHMETH.2288

Jayaprakash, A. D., Jabado O., Brown, B. D. and Sachidanandam, R. (Sept 2, 2011), Identification and remediation of biases in the activity of RNA ligases in small-RNA deep sequencing Nuc Acid Res, 1–12. doi:10.1093/nar/gkr693

Krawczynski, K., et al. (2015) Expression of microRNAs and isomiRs in the porcine endometrium: implications for gene regulation at the maternal-conceptus interface. BMC Genomics. 16:906. doi: 10.1186/s12864-015-2172-2.

Langlois, R. A. et al. (Nov 15, 2011), In Vivo Delivery of Cytoplasmic RNA Virus-derived miRNAs. Molecular Therapy. doi:10.1038/mt.2011.244

Nakashe, P. et al. (2011) Adapter Dimer Reduction in High-Throughput microRNA Profiling. OMICS 1(1): 6-11.


Kit Specs

CONCENTRATION: 200 U/uL

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.

SOURCE: Purified from an E. coli strain carrying T4 RNA Ligase 2 truncated overproducing plasmid.

STORAGE: Store at -20°C

SHELF LIFE: 1 year when stored under optimal conditions

STORAGE & DILUTION BUFFER: 10 mM Tris-HCl (pH 7.5), 100 mM NaCl, 0.1 mM EDTA, 0.1 mM DTT and 50% Glycerol

QUALITY CONTROL: Enzyme preparations are routinely assessed for relative purity, activity and absence of RNase and DNase. In addition, this enzyme is tested for reactivity with adenylated linkers (catalog # 510205, 510305, 510405)

Validated on Illumina® sequencing platforms.

Bioo Scientific’s AIR™ Ligase was developed specifically for demanding Next-Generation RNA Sequencing applications. As seen in Figure 1, AIR Ligase 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. AIR Ligase is a truncated version of T4 RNA Ligase 2. Unlike the full length ligase, AIR 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. Figure 1 illustrates that AIR Ligase is much more efficient than the competitor’s T4 RNA Ligase 2, truncated as indicated by the ligation product in lane 1. Whether you plan to sequence microRNAs or perform directional mRNA-Seq, AIR Ligase will enhance your library preparations.

AIR Ligase Ligation versus Company X's Ligase

 

Figure 1. AIR Ligase ligation of a 25-mer microRNA with a 24-mer AIR™ Adenylated Linker. Lane 1 demonstrates a ligation using AIR™ Ligase (20 U) with supplied buffer. Lane 2 is an example of a competitor’s T4 RNA Ligase 2, truncated (20 U) with supplied buffer.  Lane 3 is identical to lane 2 except that 40 U of competitor T4 RNA Ligase 2, truncated is used. These incubations were carried out at 2 hours at room temperature and run on a denaturing 15% TBE gel.