ROCHE First Strand cDNA Synthesis Kit for RT-PCR (AMV)

Specific Activity: >50 units/μg protein

Storage: −20°C

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

Application:
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

General description:
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.

Other Notes:
For life science research only. Not for use in diagnostic procedures.

Quality:
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.

Specificity:
Heat inactivation: 5 min, 95 °C
Reverse Transcriptase AMV