Specific Activity: >50 units/μg protein
UNSPSC Code: 41105903
Components: Reverse Transcriptase AMV; PCR Nucleotide Mix, pH 8.5 10 mM each; Reaction Buffer; MgCl2 Stock Solution 25 mM; Gelatin 0.05% (w/v); Oligo-p(dT)15 , 0.02 A260 units/μl 0.8 μg/μl; Random Primer p(dN)6 , 0.04 A260 units/μl 1.6 μg/μl; RNase Inhibitor 50 U/μl; Control Neo pa RNA (1.0 kb in length with additional 19-base 3′-poly(A) tail) 0.2 μg/μl; Water, PCR Grade
RIDADR: NONH for all modes of transport
The First Strand cDNA Synthesis Kit can be used with either sequence-specific primers, poly(dT)15 primers, or random primers, or gene-specific primers (not part of the kit). It can be used for the:
• Detection of the presence or absence of RNA viruses or other RNA-containing microorganisms (in combination with PCR)
• Quantification of mRNA for monitoring differential expression of a specific mRNA
• First step in the "differential display of mRNA"
• Generation of cDNA libraries with large and full-lenght inserts
The amplification of RNA requires the conversion of the RNA substrate into DNA. This is achieved through the use of a reverse transcriptase such as AMV RT (avian myeloblastis virus reverse transcriptase) or M-MuLV RT (moloney murine leukemia virus reverse transcriptase). The resulting cDNA can be used as a template for a standard PCR.
AMV RT synthesizes the new cDNA strand at site(s) determined by the type of the primer used:
• at the 3′-end of the poly(A) mRNA when Oligo-p(dT)15is used as a primer,
• at nonspecific points along the mRNA template when using the random primer p(dN)6, or
• at a site determined by a sequence-specific primer.
For life science research only. Not for use in diagnostic procedures.
In the standard cDNA synthesis assay, at least 300 ng of cDNA is synthesized (i.e., 30% yield) when 1.0 μg Neo pa RNA template is incubated with 20 μCi [α-32P]-dCTP (specific activity of 3000 Ci/mmol) for 60 minutes at +42°C.
Heat inactivation: 5 min, 95 °C
Reverse Transcriptase AMV