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ERIC Number: EJ920851
Record Type: Journal
Publication Date: 2011
Pages: 10
Abstractor: As Provided
Reference Count: 13
ISBN: N/A
ISSN: ISSN-1470-8175
Real-Time PCR (qPCR) Primer Design Using Free Online Software
Thornton, Brenda; Basu, Chhandak
Biochemistry and Molecular Biology Education, v39 n2 p145-154 Mar-Apr 2011
Real-time PCR (quantitative PCR or qPCR) has become the preferred method for validating results obtained from assays which measure gene expression profiles. The process uses reverse transcription polymerase chain reaction (RT-PCR), coupled with fluorescent chemistry, to measure variations in transcriptome levels between samples. The four most commonly used fluorescent chemistries are SYBR[R] Green dyes and TaqMan[R], Molecular Beacon or Scorpion probes. SYBR[R] Green is very simple to use and cost efficient. As SYBR[R] Green dye binds to any double-stranded DNA product, its success depends greatly on proper primer design. Many types of online primer design software are available, which can be used free of charge to design desirable SYBR[R] Green-based qPCR primers. This laboratory exercise is intended for those who have a fundamental background in PCR. It addresses the basic fluorescent chemistries of real-time PCR, the basic rules and pitfalls of primer design, and provides a step-by-step protocol for designing SYBR[R] Green-based primers with free, online software. (Contains 8 figures.)
John Wiley & Sons, Inc. Subscription Department, 111 River Street, Hoboken, NJ 07030-5774. Tel: 800-825-7550; Tel: 201-748-6645; Fax: 201-748-6021; e-mail: subinfo@wiley.com; Web site: https://secure.interscience.wiley.com/cgi-bin/jhome/112782101
Publication Type: Journal Articles; Reports - Descriptive
Education Level: Higher Education; Postsecondary Education
Audience: N/A
Language: English
Sponsor: N/A
Authoring Institution: N/A