Sensitive single-stage PCR using custom-synthesized internal controls.
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.
Zimmermann
K
K
Baxter Hyland Immuno, Vienna, Austria.
Rieger
M
M
Gross
P
P
Turecek
P L
PL
Schwarz
H P
HP
D000818:Animals; D017931:DNA Primers / Q000737:chemistry; D004266:DNA Virus Infections / Q000175:diagnosis / Q000821:virology*; D004267:DNA Viruses / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000235:genetics; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D005169:Factor VIII / Q000235:genetics; D005838:Genotype; D006525:Hepatitis, Viral, Human / Q000175:diagnosis / Q000821:virology*; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D016732:Parvovirus B19, Human / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D005169:Factor VIII