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Indhumathi

Member Since 28 May 2012
Offline Last Active May 13 2013 08:19 PM

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Active Posts 9
Profile Views 327
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Age Age Unknown
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Gender
Female
Location
Tamilnadu,India

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My research interests
Cell culture, Gene silencing, Antisense Technology, Cancer Therapy, Angiogenesis

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Topics I've Started

will mRNA from different cell line lead to loss of expected product in RT PCR?

17 April 2013 - 09:19 PM

Hi all,
I have set of primers designed for Rat mRNA and I have previously used these primers in RT PCR with mRNA isolated from RIN cells and got the amplicon of expected size. Now I am repeating the same with mRNA isolated from INS 1E and I am not able to get the expected product in RT PCR, I even optimized the annealing temperature. I am getting a non specific product of different size if the annealing temperature is changed, but no band is seen while using the previously optimized annealing temperature. I can not decipher what is just happening. Will the change in template source affect the RT PCR? both are rat cell lines only.
Please someone help me in this.
Indhu.

PCR with Plasmid recovered from filter paper

26 March 2013 - 08:59 PM

Hi,
I have got a plasmid on filter paper. After recovery, can I use it directly for PCR? I read in a blog that it should only be used for transformation.
Indhu

PCR - consistent false positive results

06 September 2012 - 11:06 PM

Hi all,
I am in the first year of my PhD and I am facing a great deal of problem with PCR. Mock PCR with sterile water also gives positive result with exact size of my product (around 120 bp). Is it possible to have such a persistent contamination problem to get same size band always. I cloned single inserts which is around 120 bp. the real problem came up when I tried to clone two inserts which should have given 250 bp band. But it also came around 120 bp even after so many repeats. Then I went for mock PCR with sterile water which also gave the same result. What could be the problem? does hairpin loop in primers cause self priming and lead to amplification in PCR?
Indhu