By Chhandak Basu
This quantity presents an outline on layout PCR primers for profitable DNA amplification. Chapters specialise in primer layout recommendations for quantitative PCR, in silico PCR primer layout, and primer layout utilizing software program. Written within the hugely profitable equipment in Molecular Biology sequence structure, chapters comprise introductions to their respective subject matters, lists of the required fabrics and reagents, step by step, without problems reproducible laboratory protocols, and tips about troubleshooting and heading off recognized pitfalls.
Authoritative and simply available, , moment version seeks to assist molecular biology scholars, researchers, professors and PCR fans.
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Additional resources for PCR Primer Design (2nd Edition)
Based on nearest-neighbor thermodynamics, OLIGO’s search algorithms find optimal primers for PCR, qPCR (TaqMan® probes), and sequencing. OLIGO 7 searches also for hybridization, ligase chain reaction probes and molecular beacons and even siRNAs . Primer3 is one of the most commonly used primer design software . It is a frequently updated, open-source project and used by many Web-based applications to develop useful functions for primer and probe design . Its popularity is likely due to several factors that include the availability of a relatively easy-to-use Web 38 Alicia Rodríguez et al.
Different dye labels can be added to the 5′ end of the primer to differentiate amplicons, even those of the same length, by the unique fluorescence . Of primary interest in forensic science is copying short tandem repeat (STR) microsatellite loci as they are highly variable and suitable for human DNA typing applications. Several commercial kits have been developed for the purpose identifying STR alleles at loci on the autosomes and sex chromosomes for use in human identification. Primers may also be designed to evaluate the base identity in single nucleotide polymorphisms (SNPs) in with much smaller PCR amplicons than STRs.
13. 14. 15. 16. an enhanced web interface to Primer3. Nucleic Acids Res 35:W71–W74 Elkins KM (2011) Designing PCR primer multiplexes in the forensic laboratory. J Chem Educ 88:1422–1427 Ruitberg CM, Reeder DJ, Butler JM (2001) STRBase: a short tandem repeat DNA database for the human identity testing community. Nucleic Acids Res 29:320–322 Papp AC, Pinsonneault JK, Cooke G, Sadée W (2003) Single nucleotide polymorphism genotyping using allele-specific PCR and fluorescence melting curves. Biotechniques 34: 1068–1072 Wang J, Chuang K, Ahluwalia M, Patel S, Umblas N, Mirel D, Higuchi R, Germer S (2005) High-throughput SNP genotyping by single-tube PCR with Tm-shift primers.
PCR Primer Design (2nd Edition) by Chhandak Basu