What is first strand cDNA synthesis?

What is first strand cDNA synthesis?

The synthesis of DNA from an RNA template, via reverse transcription, produces complementary DNA (cDNA). Alternatively, the first-strand cDNA can be made double-stranded using DNA Polymerase I and DNA Ligase. These reaction products can be used for direct cloning without amplification.

What is cDNA synthesis kit?

cDNA synthesis kits provide a fast and reliable means of producing cDNA from RNA samples. These convenient ready-to-use kits come with the components necessary for reverse transcription, including the reverse transcriptase (RT), dNTPs, reaction buffer, nuclease-free water, and RNase inhibitors.

What can be used to initiate first strand cDNA synthesis?

First-strand and second-strand cDNA synthesis The first-strand cDNA (with or without the RNA annealed to it) may be used directly in some applications such as RT-PCR, where a thermostable DNA polymerase (e.g., Taq DNA polymerase) replicates the complementary strand of cDNA.

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.

Is cDNA single or double-stranded?

In genetics, complementary DNA (cDNA) is DNA synthesized from a single-stranded RNA (e.g., messenger RNA (mRNA) or microRNA (miRNA)) template in a reaction catalyzed by the enzyme reverse transcriptase.

Why are random primers used?

Random Primers are oligodeoxyribonucleotides (mostly hexamers) used to prepare labeled DNA probes from templates for filter hybridization or in situ hybridization and to prime mRNAs with or without poly(A) for cDNA synthesis.

Why we use cDNA instead of DNA?

There are several advantages to using cDNA as opposed to genomic DNA for doing this: No introns: Eukaryote genes commonly contain introns (non-coding sequences). These are removed after mRNA synthesis so cDNA contains no introns. This means that a cDNA copy of a gene can be isolated as a single, intron-free fragment.

How is the transcriptor first strand cDNA synthesis Kit performed?

Figure 3: Performance of the Transcriptor First Strand cDNA Synthesis Kit in two-step RT-PCR. Total RNA was reverse transcribed using the anchored-oligo (dT) 18 primer and reaction conditions outlined in the pack insert of the product. In the subsequent PCR, 5 µl of the cDNA reaction was used.

What kind of primers are used in first strand cDNA synthesis?

The kit provides all reagents required for first-strand cDNA synthesis reactions. For priming, three different primer systems can be used. Two cDNA synthesis primers are provided with the kit: random hexamer primers and an anchored-oligo (dT) 18 primer.

What are the reagents in the primescript 1st strand synthesis kit?

PrimeScript 1st strand cDNA Synthesis Kit PrimeScript 1st strand cDNA Synthesis Kit The PrimeScript 1st strand cDNA Synthesis Kit contains all of the reagents necessary for synthesis of full-length first-strand cDNA from total or poly A + RNA, including both oligo (dT) and random 6-mer primers.

When to use reverse transcriptase in cDNA synthesis?

Due to its thermostability, Transcriptor Reverse Transcriptase is recommended for GC-rich templates with high secondary structure, without the need to include additives in the reaction. The kit provides all reagents required for first-strand cDNA synthesis reactions. For priming, three different primer systems can be used.