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GordonLeung

Member Since 19 Sep 2012
Offline Last Active May 12 2013 11:12 PM

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Gordon Leung
My research interests
Viral DNA detection, STR analysis, phylogenetics

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bioforum
19 Sep 2012 - 20:21

Topics I've Started

Shift in peaks at dissociation curve

03 October 2012 - 08:46 PM

Dear all,

I am trying to amplify my target gene from viral DNA for making a standard curve. I have used qPCR (Sybr system) instead of ordinary PCR to monitor the specificity. Regarding the dissocation curve, the peak corresponds to genomic DNA template has a melting temperature at ~81.5 oC.

After that, I have purified the PCR products using gel purification kit. A 10-fold serial dilution of the amplicon have been done and they are subjected to a second qPCR (with the same reagent and thermo-cycling profile as the previous one). Surprisingly, everything goes well execpt the peak at the dissociation curve has shifted to ~85 oC. Is it contamination? Or the melting temperature of my target gene has changed after purification? Attached please see the dissociation curve for that. (I'm sorry for the low resolution)

I have identified the amplicon by RFLP and it is the gene of my target.

Comments are always welcomed!

Synthesis of carrier RNA(poly-A) for DNA Extraction

19 September 2012 - 05:20 AM

Dear All,

My lab is planning to synthesize some carrier RNA (poly-A) to increase the yield of (viral) DNA precipitation. Does the length of poly-A RNA matters? What should be the best length to be synthesized? Thanks all!