Chapter title |
COLD-PCR: Applications and Advantages
|
---|---|
Chapter number | 2 |
Book title |
Clinical Applications of PCR
|
Published in |
Methods in molecular biology, February 2016
|
DOI | 10.1007/978-1-4939-3360-0_2 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3358-7, 978-1-4939-3360-0
|
Authors |
Zuo, Zhuang, Jabbar, Kausar J., Zhuang Zuo, Kausar J. Jabbar |
Editors |
Rajyalakshmi Luthra, Rajesh R. Singh, Keyur P. Patel |
Abstract |
Co-amplification at lower denaturation temperature-based polymerase chain reaction (COLD-PCR) is a single-step amplification method that results in the enhancement of both known and unknown minority alleles during PCR, irrespective of mutation type and position. This method is based on exploitation of the critical temperature, Tc, at which mutation-containing DNA is preferentially melted over wild type. COLD-PCR can be a good strategy for mutation detection in specimens with high nonneoplastic cell content, small specimens in which neoplastic cells are difficult to micro-dissect and therefore enrich, and whenever a mutation is suspected to be present but is undetectable using conventional PCR and sequencing methods. We describe in this chapter our COLD-PCR-based pyrosequencing method for KRAS mutation detection in various clinical samples using DNA extracted from either fresh or fixed paraffin-embedded tissue specimens. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Sweden | 1 | 7% |
Unknown | 14 | 93% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Bachelor | 4 | 27% |
Student > Ph. D. Student | 4 | 27% |
Professor | 1 | 7% |
Unknown | 6 | 40% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 6 | 40% |
Medicine and Dentistry | 1 | 7% |
Unknown | 8 | 53% |