Second Strand cDNA Synthesis Kit is an efficient system of generating double stranded cDNA from first strand cDNA as a template. The E. coli RNase H nicks the RNA in the DNA:RNA hybrid, while the E. coli DNA Polymerase replaces the RNA with deoxyribonucleotides. The E. coli DNA Ligase completes the double stranded DNA formation by linking the gaps between the newly synthesized cDNA strand. The dNTP based kit (Cat. No. G475) and dNTP/dUTP based kit (Cat. No. G476) provides different combination of deoxyribonucleotides tailored to the end user’s needs and application.
This kit is provided as individual enzymes to meet the customer’s needs and to provide maximum efficiency and flexibility in the RNA sample preparation. (Noted that these enzymes may still contain E. coli gDNA) The double stranded cDNA end product can subsequently be converted to blunt ended DNA fragments using abm’s DNA End Repair Kit (Cat. No. G477), followed by abm’s dA Tailing Kit (Cat. No. E009) to generate DNA suitable for whole genome sequencing.
Template
RNA
Format
Kit
Quantity
25 x 50 µl reactions
Reaction
RT-PCR
Reference
Chung, G et al. "CHL-1 provides an essential function affecting cell proliferation and chromosome stability in Caenorhabditis elegans." DNA Repair (Amst.) 10 (11):1174- 1182 (2011). DOI: 10.1016/j.dnarep.2011.09.011. PubMed: 21968058. Application: PCR.
Jones, MR et al. "The atm-1 gene is required for genome stability in Caenorhabditis elegans." Mol. Genet. Genomics 287 (4):325-335 (2012). DOI: 10.1007/s00438-012-0681-0. PubMed: 22350747. Application: PCR.
Lee, G et al. "Potential monoclonal antibody therapy for the treatment of ovarian cancer." Ovarian cancer—basic science perspective :385-406 (2012). DOI: 10.5772/1268 . Application: Reverse Transcription.
Unsoy, G et al. "Chitosan magnetic nanoparticles for pH responsive Bortezomib release in cancer therapy." Biomedicine & Pharmacotherapy 68(5):641-649 (2014). DOI: 10.1016/j.biopha.2014.04.003. PubMed: 24880680. Application: Reverse Transcription.
Storage Condition
-20C
Shipping Condition
Dry Ice
Storage
Store all components at -20°C. All components are stable for 1 year from the date of shipping when stored and handled properly. Avoid repeated freeze-thaw cycles to retain maximum performance.
Reviews of Second-Strand cDNA Synthesis Kit-dNTP/dUTP based