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Sensitive Single-Stage PCR Using Custom-Synthesized Internal Controls
Author(s) -
Klaus F. Zimmermann,
Miriam Rieger,
Patricia Gross,
P.L. Turecek,
Hans Peter Schwarz
Publication year - 2000
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/00284st05
Subject(s) - genotyping , oligonucleotide , microbiology and biotechnology , virus , biology , lysis , dna , virology , polymerase chain reaction , chemistry , genetics , genotype , gene
A new approach for an internally controlled PCR was developed using a custom-synthesized oligonucleotide as the internal control. Three different PCR setups demonstrated the usefulness of this approach: (i) the addition of the respective internal control to samples containing ssDNA virus Parvo B19; (ii) the co-extraction of plasma samples and the respective internal control for the detection of the ssDNA virus TTV; and (iii) the addition of the respective internal control to a crude lysate of tail pieces for the genotyping of FVIII knockout mice, demonstrating that this approach is also applicable for dsDNA.

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