What is 5 RACE PCR?

What is 5 RACE PCR?

What is 5 RACE PCR?

The 5′ RACE System is a set of prequalified reagents intended for synthesis of first strand cDNA, purification of first strand products, homopolymeric tailing, and preparation of target cDNA for subsequent amplification by PCR. Control RNA, DNA, and primers are provided for monitoring system performance.

What is RACE RNA?

Rapid Amplification of cDNA Ends (RACE) is a procedure for amplification of nucleic acid sequences from a messenger RNA template between a defined internal site and unknown sequences at either the 3′ or the 5′ -end of the mRNA (1).

What do you mean by race What are the applications of 5 and 3 race?

3´ RACE System for Rapid Amplification of cDNA Ends › 5´ RACE System for Rapid Amplification of cDNA Ends › cDNA Synthesis Directly from Cells Using SuperScript III CellsDirect cDNA Synthesis System › cDNA Synthesis from Transcripts with High Secondary Structure Using Thermo-X Reverse Transcriptase ›

What is the 5′ race system?

The 5′ RACE System provides a set of prequalified reagents intended for synthesis of first-strand cDNA, purification of first-strand products, homopolymeric tailing, and preparation of target cDNA for subsequent amplification by PCR. Control RNA, DNA, and primers are provided for monitoring system performance.

How do you electrophoresis 5′ RACE products?

9. Analyze 5-20 µl of 5′ RACE products by agarose gel electrophoresis according to standard protocols, using appropriate size standards (19). Either TAE or TBE electrophoresis buffer may be used for the procedure. The volume of the sam- ple used for analysis will dependent on the volume and thickness of the sample well.

How to analyze 5′ RACE products?

Analyze 5-20 µl of 5′ RACE products by agarose gel electrophoresis according to standard protocols, using appropriate size standards (19). Either TAE or TBE electrophoresis buffer may be used for the procedure.

What primers are required for 5′ race?

A minimum of two antisense gene-specific primers (GSP) are required for 5′ RACE and must be supplied by the user. GSP1 primes first-strand cDNA synthesis. A second nested primer, GSP2, that anneals to sequences located 3′ (with respect to cDNA not mRNA) of GSP1 is required for PCR.

What is race technique?

Rapid amplification of cDNA ends (RACE) is a technique used in molecular biology to obtain the full length sequence of an RNA transcript found within a cell.

Why do we race PCR?

RACE (rapid amplification of cDNA ends) PCR is useful for quickly obtaining full length cDNAs for mRNAs for which only part of the sequence is known and to identify alternative 5′ or 3′ ends of fully sequenced genes.

What is the second primer in the case of 5 race?

During PCR, the thermostable DNA polymerase is directed to the appropriate target RNA by a single primer derived from the region of known sequence; the second primer required for PCR is complementary to a general feature of the target-in the case of 5′ RACE, to a homopolymeric tail added (via terminal transferase) to …

What is difference between DNA and cDNA?

The main difference between DNA and cDNA is that DNA is composed of both coding and non-coding sequences whereas cDNA only contains the coding sequences. The coding sequences are the exons of a gene, which codes for a functional protein. The non-coding sequences are the remaining DNA sequences of the genome.

What is the difference between cDNA and genome DNA?

The main difference between cDNA and genomic DNA is that cDNA represents the transcriptome of a particular organism whereas genomic DNA represents the genome.

What is AFLP marker?

Summary. Amplified fragment length polymorphism (AFLP) is a PCR-based technique that uses selective amplification of a subset of digested DNA fragments to generate and compare unique fingerprints for genomes of interest.

What is a quencher in PCR?

The quencher molecule quenches the fluorescence emitted by the fluorophore when excited by the cycler’s light source via Förster resonance energy transfer (FRET). As long as the fluorophore and the quencher are in proximity, quenching inhibits any fluorescence signals.

How long can cDNA be stored at?

If the sample must be stored for a longer time, add 0.5 µl of 10X NEBNext Cell Lysis Buffer to the cDNA (after RT instead of during the PCR step). The samples can then be stored at -20˚C for up to 7 days, but a slight decrease in yield may be observed.