How to take DNA samples from snakes?

Originally Posted by bcherps

Hey,

After a talk with the boss, and another professor across the hall I have determined that Mitochondrial DNA I either was playing hooky or my eyes were glazed over from all the excitement of the lecture.

I guess it was about time for my one annual post were I talk out of my bum and someone catches me on it. Well, this made me put out way to much effort and my head hurts now-off to gather some adult beverages and wash some rubbermaids.

Thanks for the discussion though, it made me get the brain cells going! I was a lab tech / research assistant in this field once upon a time, but it was a LONG time ago, and my own knowledge is severely out of date. It's an interesting area, thanks for the replies and all the details!
Helen

Hi,
I rarely visit this forum but was informed that this post was here and that I was mentioned.
Here is my take on DNA quality and testing in reptiles.
I have been involved with DNA studies of a number of wild populations (Hog Islands, and the Boas of Cozumel). My involvment was initially developing molecular methods for screening the snakes of these populations for heterozygosity and genetic diversity, hence we could then estimate measures such as inbreeding, relatedness, effective population size, bottlenecks.founder events, etc. At the time I was developing DNA fingerprinting markers (microsatellites) for a number of other species, and given that I maintain a relatively large collection of boas myself (~100), I figured it would be relatively easy to ass these to the workload.
So, I extracted DNA from shed skins using a number of methods, namely the Puregene and Qiagen extraction kits, and also a modified phenol/chloroform method. All methid yielded extremely high quality DNA (high molecular weight with little to not degradation). I used this to develop a boa constrictor specific set of markers, and then blasted the sequences in Genbank to determine if we had homolgies or contamination. No contamination was detected, and I screened my collection for polymorphism at each locus. These markers proved viable for individual identification. Since then I ave used them in my own collection to determine the paternity of litters when multiple males were used. I am also using this class of marker in a parthenogenesis study of squamate reptiles currently, with the first study due to be published in the next few months.
So, to answer the question. Blood is ideal, if stored correctly. Tissue is also just as good. Failing that, shed skin, will yield sufficietly high quality DNA is stored dry in individual ziplock bags or envelopes. I routinely extract DNA from approximately 2 cm of shed skin.
If the individual with the potentially parthenogenetic gecko is still requiring testing, I can perform this analysis.

Warren Booth

I sent a link to the guy with the gecko.

He hasn't been on the forum in a while, but when he does come back, the message is there for him.

Thanks so much for your help!!!

Many thanks Warren, that is VERY helpful information.