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Frog Egg Physilogy

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Frog Egg Physilogy

Postby Swordy » Tue Apr 21, 2009 9:22 pm

Hi! In one of my Research Projects i have been stuck with the Troublesome task of extracting DNA from some Unfertilized frog eggs… they are Chiricahua Leopard Frog (Rana chiricahuensis) eggs and I have asked many “experts” on this type frog and they knew nothing about them in the slightest. As Such I was wondering what is known about related frogs? Do the Nucleases have a Lipid body around them? Is there a protein in frog eggs that binds to frog DNA and with prevent TAQ from working in PCR?
Anything you happen to know could prove useful as I am at my wits end as to what to do with the dang things! or if you have an Extraction Method for Frog eggs i would be eternally greatful :D

Thanks!

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Postby MrMistery » Thu Apr 23, 2009 1:54 pm

Well i would think if you wanna do PCR you can just put them in distilled water and the put them in the PCR reaction.
Anyway, i think that you would have better chance looking up a protocol for Xenopus laevis, people have been studying the eggs of that organism for decades. Then try that and see if it works
"As a biologist, I firmly believe that when you're dead, you're dead. Except for what you live behind in history. That's the only afterlife" - J. Craig Venter
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Re: Frog Egg Physilogy

Postby Swordy » Thu Apr 23, 2009 5:51 pm

Thanks for the Other Frog to look at Mr. misery :) i found some potentially usefull info on that end.

On the PCR side of things i am using unfirtilized Frog eggs meaning in theory there should be only 1 set of nuclear DNA chromosomes on the Egg masses which means i have to do everything inside of a single Sterile PCR tube without moving anything or i leave the DNA behind.

If it helps here is a list of what i have tryed already...
I have tryed simply putting the Egg inside of a PCR tube with Water, Taq, and the Forward and reverce primers with no success. I have also used various extraction DNA extraction Kits such as the MoBio Clean DNA kit, A Zippy Fecal kit and many others. I have also tryed some homegrown methods involveing in different combinations Bead Bashing, Protein Kinase, SDS, HCl, NaOH, Tris, Freezing, and Boiling. None of these have worked in the slightest except to get mtDNA which as far as i know is useless for finding out if 2 frogs are related which is the overall goal of the project.

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