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STANDARDISATION OF MULTIPLEX PCR


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#1 THIRUMURUGAN

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Posted 25 May 2011 - 11:12 PM

we like to standardise a multiplex PCR for BACTERIA, P.acne, Fungal from a clinical samples. when I did Multiplex PCR am getting only fungal and p.acne POSITIVE but negative for bacteria, but when the same condition and reaction in UNIPLEX it gives good result for all. what i should do now???

and i like to know whether 15mM Mgcl2 which is am using it.But literature says for that use 5mM Mgcl2. Whether this may inhibit the multiplex or not??


waiting for your valuable suggestion.

#2 Adrian K

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Posted 26 May 2011 - 01:12 PM

hmn.. had the paper been published before? you design the primers your own?
Expecting the world to treat you fairly because you are a good person is like expecting the lion not to attack you because you are a vegetarian.

..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...

"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong

"It's all just DNA. Do it."---phage434

#3 THIRUMURUGAN

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Posted 15 June 2011 - 12:04 AM

hmn.. had the paper been published before? you design the primers your own?

yes sir, its already published paper only. i just followed all as such in paper.

#4 Adrian K

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Posted 15 June 2011 - 01:13 AM

okie, did you perform a gradient PCR? this is because temperature varies from machine to machine... although it may published 55C...but you might need push up further to 58C...

can you show us the paper? any links?
Expecting the world to treat you fairly because you are a good person is like expecting the lion not to attack you because you are a vegetarian.

..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...

"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong

"It's all just DNA. Do it."---phage434




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