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optimal template dna concentrations


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#1 GradMicro

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Posted 06 September 2009 - 05:36 AM

Hi,
I am extracting bacterial DNA from human blood using qiagen mini amp kits. Yield seems low when calculated ussing spectrometry.

Would be grateful if anyone could tell me
1. a better extraction kit (low cost as possible) or a method (my organisms is intracellular!!)

2. the optimal range of template to be put in a 25 micro L reaction mix

3. the maximun amout of dna template volume possible to be put in a reaction mix of 25 micro L reaction mix

4. For a validation of a PCR the best dilusion series for a positive control to get the minimum detection limit (conc of my positive control is 390 microgrms per ml

#2 Adrian K

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Posted 09 September 2009 - 10:30 PM

Try use Promega genomic purification kits.
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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