i am working whit philogenetics whit mitochondrial DNA dloop sequencing on poultry. I am using a rapid DNA extraction from whoole blood whit two methods:
1. wash blood whit TE buffer 2 time
2. digesting whit proteinase K and PCR buffer
1) 1ul of whoole blood in 300 5% chelex resin
2) incubated 1 hour at 60 C and 15 min at 95
Both methods work excellently for microsatellites analisis but when i used this DNA for mitchondrial DNA I did obtain any amplification. I use a positive control extracted whit phenol-croloformium mehtod and I obtained strog amplification both from microsatellites locus as mitochondrial fragments.
So my question is...could rapid methods be not enogh to lyse mitochondrial membrane?
Edited by vincio, 17 March 2012 - 03:32 AM.