Analysis Note

To view the Certificate of Analysis for this product, please visit www.cytiva.com.

Application

First-strand cDNA synthesis kit has been used to reverse transcribe total RNA into cDNA.

Features and Benefits

For synthesis of single-stranded cDNA prior to amplification by PCR. Features preassembled bulk reaction mixes to save time and minimize potential for pipetting errors. Allows users to choose primers. Includes a choice of two primers: pd(N)6 random hexamers and NotI-(dT)18.

General description

Designed to generate full-length first-strand cDNA from mRNA templates. Following first-strand cDNA synthesis, the sample can be used directly for in vitro amplification using PCR or for second-strand synthesis using the Gubler-Hoffman method.First-Strand cDNA Synthesis Kit is designed to generate full-length first-strand cDNA from mRNA templates. Following first-strand cDNA synthesis, the sample can be used directly for in vitro amplification using PCR or for second-strand synthesis using the Gubler-Hoffman method. Primers are not included in the preassembled First-Strand Reaction Mixes, thus allowing use of a primer of choice. Two primers are included with the kit. pd(N)6 Primer (random hexamer) is used to prepare libraries for screening with antibodies, to increase the representation of 5′-ends of mRNAs having significant secondary structure, or to copy mRNAs lacking a poly(A) tail.The NotI-(dT)18 Bifunctional Primer can be used to prime selectively on mRNA that has a poly(A) tail. It can also be used as a PCR primer following first-strand synthesis. The NotI restriction site, located at the 5′-end of the primer, is particularly useful for subsequent directional cloning of cDNA and PCR products.First-Strand cDNA Synthesis Kit contains the following reagents, sufficient for up to 55 first-strand syntheses, each of which will produce enough material for up to 15 amplification reactions: Bulk First-Strand cDNA Reaction Mixes, mRNA Standard, DTT Solution, RNase-Free Water, pd(N)6 Primer, NotI-(dT)18 Bifunctional Primer, and instruction booklet.