Protocol Online logo
Top : New Forum Archives (2009-): : PCR, RT-PCR and Real-Time PCR

optimal template dna concentrations - (Sep/06/2009 )

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


Try use Promega genomic purification kits.

-adrian kohsf-