A novel and rapid method for synthesizing positive controls and standards for quantitative PCR.
We developed and tested a method to produce DNA standards and controls for quantitative PCR by designing and performing partial hybridization of long oligonucleotides before double stranded DNA fragments were synthesized and subsequently amplified by conventional PCR. This approach does not require any natural DNA template. Applications include the production of standards, which cannot be easily produced from DNA extracted from bacteria or plants.
David
Maude M
MM
Environmental Microbial Genomics Group, Microsystems and Microbiology, Laboratoire Ampère, UMR CNRS 5005, Ecole Centrale de Lyon, Université de Lyon, 36 avenue Guy de Collongue 69134 Ecully cedex, France.
Sapkota
Amy R
AR
Simonet
Pascal
P
Vogel
Timothy M
TM
D002138:Calibration; D017931:DNA Primers / Q000138:chemical synthesis* / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards*; D012015:Reference Standards; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004269:DNA, Bacterial