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closeUPDATE: use of synthetic gene fragments as Bd qPCR standards
Posted by analongo on 12 Nov 2013 at 21:22 GMT
Several companies are synthesizing small oligo fragments that can be used as standards for qPCR. We recently ordered Haplotype 1 (see Appendix S1) from IDTDNA (gBlocks Gene Fragment), which is supplied dry in 200ng. The gBlock gene fragments are a good alternative to the PCR fragment quantification method that we discussed in this paper, because they eliminate the need of PCR amplification and DNA quantification, both steps that potentially introduce error. As a reference for other researchers performing these qPCR experiments with gBlocks as the standard, our serial dilution curve starting with 10^6 gBlock fragments amplified at an average cycle threshold of 21.92. The authors recommend the use of these fragments to quantify ITS copies from different Bd strains as the most comparable method for Bd infection load quantification across laboratories and strains.