What is cDNA synthesis kit?

Thermo Scientific RevertAid First Strand cDNA Synthesis Kit is a complete system for efficient synthesis of first strand cDNA from RNA templates. The kit uses RevertAid Reverse Transcriptase (RT), a recombinant M-MuLV RT which maintains activity at 42-50°C and is suitable for synthesis of cDNA up to 13 kb.

What is the purpose of cDNA synthesis?

The synthesis of DNA from an RNA template, via reverse transcription, results in complementary DNA (cDNA). cDNA can then serve as template in a variety of downstream applications for RNA studies such as gene expression; therefore, cDNA synthesis is the first step for many protocols in molecular biology.

What does reverse transcriptase do?

Reverse transcriptases have been identified in many organisms, including viruses, bacteria, animals, and plants. In these organisms, the general role of reverse transcriptase is to convert RNA sequences to cDNA sequences that are capable of inserting into different areas of the genome.

What’s the protocol for Invitrogen real time PCR?

Add: 1 ul RNase H, mix, incubate at 37°C, 20 min, 95°C, 10 min (an additional step to the Invitrogen protocol to remove any components that may interfere with the PCR (Ambion website). Store cDNA at -20ºC Real Time-PCR We typically dilute the cDNA reaction 1:10 with DEPC water and use 5μl as template for a 20μl RT-PCR reaction. 1.

How is superscript Vilo cDNA synthesis kit used?

The Invitrogen™SuperScript™VILO™cDNA Synthesis Kit provides the high temperature capability of SuperScript™III Reverse Transcriptase in an optimized format for generating first-strand cDNA for use in real-time quantitative RT–PCR (qRT–PCR).

What’s the protocol for qRT-PCR and cDNA synthesis?

Protocol for cDNA synthesis and qRT-PCR cDNA Synthesis From Kyle Harris and Alison Nairn: 4/12/07 Protocol for cDNA synthesis and qRT-PCR cDNA Synthesis Superscript III 1stStrand Synthesis Kit (Invitrogen, Cat #18080-051) We typically use 1μg of Total RNA per 20μl cDNA reaction, but you may be able to use less.

How much RNA is in a cDNA synthesis kit?

cDNA Synthesis Superscript III 1stStrand Synthesis Kit (Invitrogen, Cat #18080-051) We typically use 1μg of Total RNA per 20μl cDNA reaction, but you may be able to use less. Initial Primer/RNA Mix: 0.5 μl random hexamer primers 0.5 μl oligo(dT) primers 1μl dNTP mix 8ul of RNA (1μg) + DEPC H 20 Heat to 95°C, 5 min