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Questions related from Lee Lee
My project is about detection of species DNA in feed sample. I get a strange melt curve result from a developed species detection assay. The assay was optimized with genomic DNA. Peak at Tm=74 is...
17 March 2016 8,818 4 View
Hi there, I have design a 20bp forward primer which located at 2 nucleotides before TaqMan probe. The question is that, is spacing of 2 nucleotides enough for polymerase to sit on 3' end of...
14 March 2016 4,737 4 View
I have designed a TaqMan probe with 30 nt in size targeted a segment in mitochondria gene, which is conserved in most vertebrate. Multiple sequence alignment was done. There are still few...
08 March 2016 9,318 4 View
Dear all, I am currently validating an optimized duplex qpcr assay with various samples. The target primers and internal control primers specificity was tested and no cross reaction was detected....
07 March 2016 756 6 View
Many reported SYBR green multiplex PCR assays gave few significant peak within the melting curve. I have designed 2 primer sets for two different target in my project. 500nM primers were used for...
12 February 2016 6,029 8 View
The qPCR assay worked well previously with amplification efficiency of 1.901. Thing goes wrong in recent. I made a standard curve with 10-fold serial dilution gDNA as template. Late Ct value was...
21 January 2016 6,237 4 View
I have been troubleshooting the qPCR standard curve for weeks. A serial of 10-fold dilutions of porcine gDNA ( 10 2 - 10-4 ng) were tested using LightCycler 480. The run consist of 40 cycles of...
15 October 2015 9,395 8 View
