cDNA synthesis using superscript II ..

Hi,

I am trying to perform qPCR on cDNA samples from mice. I performed my cDNA synthesis with Superscript Vilo (Invitrogen), which is based on random priming. I have some samples in which I observe non-sigmoidal amplification plots, these samples have very low Ct counts (See attachment). And if I run these samples on an 1% agarose gel I observe smearing in addition to the PCR product of expected size (See attachment). For my qPCR I used 1:10 diluted cDNA with Quantitect SYBR Green Kit (Qiagen).

What are these smears on the gel, and how can I get rid of them?
Any ideas or suggestions or solutions will be greatly appreciated

THANK YOU!!





Bio-Rad's iScript cDNA Synthesis Kit | Biocompare …

03/01/2006 · A review of the Bio-Rad's iScript cDNA Synthesis Kit
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cDNA Synthesis Kits; SuperScript™ IV VILO ..

This protocol has several roots. It is based on a protocol to create cDNA for microarrays and adapted to be used for lower amounts. It works very reliable and it is my preferred way of cDNA synthesis.

RT reaction using Superscript VILO cDNA synthesis kit

This protocol describes how to make cDNA from RNA using Invitrogen's ViLo kit. This kit will make single stranded cDNA using first-strand synthesis from single stranded RNA, normally purified using Trizol. The Vilo kit uses random primers (not oligo DT).

This protocol describes how to make cDNA from RNA using Invitrogen's ViLo kit
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using the SuperScript VILO cDNA Synthesis Kit ..

We recommend using the SuperScript® VILO cDNA synthesis kit ..

SuperScript VILO cDNA Synthesis kit ..