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- 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 Not I-(dT)18.
Technical Information
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 Not I-(dT)18 Bifunctional Primer can be used to prime selectively on mRNA having a poly(A) tail. It can also be used as a PCR primer following first-strand synthesis. The Not I 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, Not I-(dT)18 Bifunctional Primer and instruction booklet.
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