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Questions related from Lee Lee
Limit of detection (LOD) and sensitivity were regards as synonymous in most of the QPCR publication works in the past. However, it has been controversially discussed as: (a) LOD refers to the...
02 February 2018 9,874 1 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...
03 March 2016 9,114 4 View
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...
03 March 2016 2,920 5 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...
01 January 2016 2,610 5 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...
10 October 2015 1,168 9 View
Dear all, I am following a gDNA rapid salt-extraction method for PCR-based techniques reported in Nucleic Acid Research Journal. Here is the protocol: 100mg minced meat homogenized in 440 µL TNES...
10 October 2015 3,274 4 View