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Second-Strand cDNA Synthesis Kit-dNTP/dUTP based

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Second-Strand cDNA Synthesis Kit-dNTP/dUTP based

品牌：Abmgood

貨號：G476

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Second-Strand cDNA Synthesis Kit-dNTP/dUTP based

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	Print Version
Description	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.
Literature
Application	RNA-Seq Library Construction. downstream double-stranded blunt-end cDNA synthesis for cloning. downstream double-stranded cDNA library construction.
Protocol
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.
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.