title: Real-time PCR-based genotyping from whole blood using Taq DNA polymerase and a buffer supplemented with 1,2-propanediol and trehalose.
abstract: Amplification of DNA templates from whole blood with Taq DNA polymerase remains a difficult task worldwide. Using a real-time PCR setup and a buffer supplemented with 1M 1,2-propanediol, 0.2M trehalose, and SYBR green I we show a reliable technique for genotyping in mice and detection of single-nucleotide polymorphisms/mutations in humans. Elimination of DNA extraction and use of the common Taq DNA polymerase and DNA dye bring about substantial savings in labor and cost.
authors.affiliation: Department of Signal Transduction, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, CZ-14220 Prague 4, Czech Republic.
authors.forename: Pavol
authors.initials: P
authors.lastname: Utekal
authors.affiliation: Department of Signal Transduction, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, CZ-14220 Prague 4, Czech Republic.
authors.forename: Lukas
authors.initials: L
authors.lastname: Kocanda
authors.affiliation: Department of Clinical Biochemistry, Regional Hospital Liberec, CZ-46063 Liberec 1, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Matousek
authors.affiliation: Department of Clinical Biochemistry, Regional Hospital Liberec, CZ-46063 Liberec 1, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Wagner
authors.affiliation: Department of Signal Transduction, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, CZ-14220 Prague 4, Czech Republic.
authors.forename: Viktor
authors.initials: V
authors.lastname: Bugajev
authors.affiliation: Department of Signal Transduction, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, CZ-14220 Prague 4, Czech Republic. Electronic address: draberpe@img.cas.cz.
authors.forename: Petr
authors.initials: P
authors.lastname: Draber
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D002021:Buffers; D004247:DNA / Q000737:chemistry; D003959:Diamines; D005838:Genotype; D006801:Humans; D051379:Mice; D009930:Organic Chemicals / Q000737:chemistry; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D019946:Propylene Glycol / Q000737:chemistry*; D011804:Quinolines; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D019914:Taq Polymerase / Q000737:chemistry*; D014199:Trehalose / Q000737:chemistry*
chemical_list: D052160:Benzothiazoles; D002021:Buffers; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D019946:Propylene Glycol; D004247:DNA; D014199:Trehalose; D019914:Taq Polymerase
keywords: 1,2-Propanediol; Real-time PCR; SYBR green I; Taq DNA polymerase; Trehalose; Unseparated blood