Product Description

First-strand cDNA Synthesis Kit 25 Reaction Kit.

CONTENTS:

This kit contains sufficient reagents to perform 25 first strand cDNA syntheses with up to 2 mg of purified poly A(+)mRNA or 10 mg of total cell RNA per reaction.

STORAGE:

12 months at -20°C
12 months at -70°C

DESCRIPTION:

This First-strand cDNA Synthesis kit has been developed to maximize the synthesis of full-length cDNA. This kit produces an optimal template for DNA amplification techniques. Tested against a 7.5 kb synthetic poly A(+)RNA and a 9.8 kb native viral mRNA, 32% and 42% of the cDNAs synthesized were full-length, respectively. Also, cDNAs to a poly A(+)RNA ladder consisted of approximately 70% full length product (Fig. 1). The use of full-length cDNA templates for polymerase chain reaction (PCR) (1) affords equal availability of all sequences for priming and amplification regardless of location. This kit includes RNase inhibitor, DTT solution, 5X reaction buffer containing nucleotides, and AMV reverse transcriptase enzyme. Pd(T)12-18 and random hexamer primers are also provided, together with RNase-free water and a polyadenylated RNA ladder as control template.

cDNA SYNTHESIS

Figure 1. [32 P]-labeled cDNA to three RNA templates:

Lanes 1 and 2: 7.5 kb synthetic poly A(+)RNA primed with pd(T)12-18 and random hexamers, respectively.
Lanes 3 and 4: 9.8 kb tobacco vein mottling virus (TVMV) mRNA, primed with pd(T)12-18 and random hexamers, respectively.
Lanes 5 and 6: poly A(+)RNA ladder* primed with pd(T)12-18 and random hexamers, respectively.

*This polyA(+) RNA ladder is different from the one supplied in this kit.

FOR LABORATORY USE ONLY

Download details and protocol.