cDNA Kit and T7T Kit

CATALOG NUMBER: LSK-1200

AMV Reverse Transcriptase
First-Strand cDNA Synthesis Kit

To download details and protocol in Adobe PDF format:

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:

• 3 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:

1. Lanes 1 and 2:                   7.5 kb synthetic poly A(+)RNA primed with pd(T)_12-18 and random hexamers, respectively.
2. Lanes 3 and 4:                   9.8 kb tobacco vein mottling virus (TVMV) mRNA, primed with pd(T)_12-18 and random hexamers, respectively.
3. 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

AdvanceScript T7 Transcription Kit

DESCRIPTION:

The AdvanceScript T7 Transcription Kit is designed for in vitro production of RNA molecules using DNA templates containing T7 promoter sequences.  The kit will be a valuable tool in a variety of applications that require generating radioactive- or non-radioactive RNA and non-labeled RNA fragments.  The kit contains all reagents sufficient for 25 or 50 reactions in a 50 µl volume of reaction mixture, and can yield 7-14 µg of RNA per 1 µg DNA template.