We are doing chelex extraction of human DNA from buccal cells. Before proceeding to PCR, I decided it would be a good idea to confirm extraction by running the genomic DNA through gel electrophoresis then stain with ethidium bromide.
While the DNA ladders show up fine, the genomic DNA is not showing up.
Is there something about chelex extraction that prevents the genomic DNA from staining?
Nanodrop analysis of the samples indicate DNA concentrations of 13- 40 ug/ul. The 260:280 ratios arent great (1.5 range) but that isnt surprising given all the the cellular material present.
Thanks in advance
Two things I can think of:
1. Full chromosomes are to large to run on a regular gel, unless you use pulse-field gel all your DNA will be stuck in the well.
2. If the DNA is sheared than it may be single strand because of the boiling step in chelex extraction. In this case you can try staining your gel with a dye that stain ssDNA like GelStar.
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining