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  2. → Viewing Profile: Topics: athalianaovule

athalianaovule

Member Since 04 Jun 2012
Offline Last Active Jul 19 2012 03:01 AM
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Topics I've Started

Genotyping genomic DNA before DNase treatment?

05 July 2012 - 09:45 AM

Hi all,

Does anybody know if I can genotype/perform genomic DNA PCR amplification with genomic primers on an RNA extraction before performing a DNase treatment? I am using the Qiagen Plant Mini RNA Prep kit, starting with a little under 100mg of tissue.  Would there be enough whole DNA within the pre-DNase treated sample to perform a genomic DNA PCR?  

I need to both genotype my plants as well as perform an RNA extraction for RT-PCR - my plants are stunted and I can not take a leaf sample prior to performing an RNA extraction for a separate DNA extraction followed by PCR.

Any feedback would be helpful.

Primer Design Help: GFP primers for Arabidopsis

13 June 2012 - 08:07 AM

Hi, I am new here, and I would like some help/advice in designing primers for GFP in Arabidopsis GFP marker lines. I want to design a "universal" primer that would recognize the GFP sequence so that I can PCR genotype my plants that have been crossed with the marker. I know that I can technically use a selectable marker (kanamycin or basta) to identify the plants that are positive for GFP - but I would rather PCR genotype for the marker and my fellow lab mates have expressed interest in using the primers I should be designing for their own work. I've looked at several papers (that deal with PCR genotyping for GFP marker lines in Arabidopsis) that have provided the GFP primer sequence that was used - but there seems to be no consensus sequence between the papers and I am now confused as to how to go about designing primers for just GFP. Any feedback would be appreciated.

Designing GFP primers for Arabidopsis

04 June 2012 - 04:47 PM

Hi, I am new here, and I would like some help/advice in designing primers for GFP in Arabidopsis GFP marker lines.  I want to design a "universal" primer that would recognize the GFP sequence so that I can PCR genotype my plants that have been crossed with the marker. I know that I can technically use a selectable marker (kanamycin or basta) to identify the plants that are positive for GFP - but I would rather PCR genotype for the marker and my fellow lab mates have expressed interest in using the primers I should be designing for their own work.  I've looked at several papers (that deal with PCR genotyping for GFP marker lines in Arabidopsis) that have provided the GFP primer sequence that was used - but there seems to be no consensus sequence between the papers and I am now confused as to how to go about designing primers for just GFP.  Any feedback would be appreciated.

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