Product Description
Biocrede’s 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 and the dNTP/dUTP based kit offer different combinations of deoxyribonucleotides tailored to the end user’s needs and application.
Application
- RNA-Seq Library Construction.
- downstream double-stranded blunt-end cDNA synthesis for cloning.
- downstream double-stranded cDNA library construction.
Protocol 
Notes
- one enzyme unit is defined as the amount of enzyme required to incorporate 1 nmol of deoxynucleotide into acid-precipitable material in 10 minutes at 37oC using Poly(A) and Oligo(dT) as template and primer, respectively.
- Noted that these enzymes may still contain E. coli gDNA.
- Reaction Buffer Components: 20 mM Tris-HCl (pH 7.5), 12 mM (NH4)2SO4, 10 mM MgCl2, 0.16 mM β-NAD.
Related Items
hiGC-2ndStructure 2X qPCR MasterMix
$125.00
Product Description
Amplify difficult DNA targets in real-time PCR with Biocrede’s hiGC-2ndStructure 2X qPCR MasterMix. Biocrede’s hiGC-2ndStructure 2X qPCR MasterMix is designed for amplification of complex DNA fragments (i.e. high GC content and secondary structure) ranging from 300bp to 1.3kb in length. Available with the option of ROX or fluorescein as the internal passive reference dye, Biocrede’s hiGC-2ndStructure 2X qPCR MasterMix enables reliable amplification and offers unmatched performance in sensitivity and signal-to-noise ratio.
Key Features
- Enable streamlined protocol in a simple reaction set-up
- Allow accurate quantification of a variety of gene targets
- Reduce pipetting steps to minimize the risk of contamination
- Compatible with most real-time PCR instruments
Application
- Next generation sequencing
- Gene-expression analysis
- Gene knockdown validation
- SNP genotyping assays
- CHiP
- Copy number variation
- Microarray validation
- High throughput applications
- Virus detection and quantification
Biocrede's hiGC-2ndStructure 2X qPCR MasterMix is supplied as 4 x 1.25ml Aliquots
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