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The Reverse Transcription System(a,b) provides reagents to efficiently reverse transcribe RNA into cDNA in fifteen minutes. The cDNA prepared from each reaction using this system may be used directly in multiple PCR reactions using Promega Taq DNA Polymerase. The AMV Reverse Transcriptase synthesizes single-stranded cDNA from total or poly(A)+ RNA. Both Oligo(dT)15 and Random Primers are included, allowing cDNA synthesis from virtually any RNA source. The system contains sufficient reagents for 100 cDNA synthesis reactions, processing 1μg of RNA per reaction. Each cDNA synthesis reaction may be divided and used in up to 20 separate PCR reactions. A polyadenylated 1.2kb RNA transcript is provided as a control template for the cDNA synthesis reaction.
Two-step amplification products generated using Primer Pairs for BDNF, CNTF, NGF, b-Actin and Promega's Reverse Transcription System.
Rat brain total RNA (1µg) was used as template; the annealing temperature during the amplification reaction was 65°C and the amplification reaction was as recommended for the Primer Pairs. Fragments were resolved on a 3.0% NuSieve® agarose gel stained with ethidium bromide. Markers are pGEM®(c) DNA Markers (Cat.# G1741); lanes with (+) are positive test reactions and lanes with (–) are negative control (no template) reactions. Haak-Frendscho, M., et al. (1997) Primer pairs for neurotophic factor mRNA analysis. Promega Notes 63, 10-12.
(a)The PCR process is covered by patents issued and applicable in certain countries. Promega does not encourage or support the unauthorized or unlicensed use of the PCR process. Use of this product is recommended for persons that either have a license to perform PCR or are not required to obtain a license.
(c)U.S. Pat.No. 5,552,302 has been issued to Promega Corporation for the methods and compositions for production of human recombinant placental ribonuclease inhibitor (PRI). Inhibitors of Angiogenin, which comprise a segment of human PRI, is the subject of U.S. Pat. Nos. 4,966,964, 5,019,556 and 5,266,687 assigned to President and Fellows of Harvard College and exclusively licensed to Promega Corporation.
(c)U.S. Pat. No. 4,766,072 has been issued to Promega Corporation for transcription vectors having two different bacteriophage RNA polymerase promoter sequences separated by a series of unique restriction sites into which foreign DNA can be inserted.
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Reverse Transcription
System
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