Damselfly DNA Extraction

[Taylor R.]

My science class at school is trying to extract DNA from damselfly larvae using an Instagene Matrix, PCR, and electrophoresis. It didn't work out the first time, so we're looking for any tips that people in the DIY community might have. Any ideas?

[Mega]

Did you use the right primers for amplifying the piece of DNA you want?

Or do you just want to see the total DNA? Then a PCR is the wrong thing to do...


Did you forget ethidium bromide in the gel?

[Cory Tobin]

Assuming you added all of the correct ingredients and you know your
primers work, I have two suggestions

1) Make sure your pipettes are calibrated and that you pipette
accurately. When I run bio classes for beginners this seems to be one
of the biggest hurdles to getting PCRs to work. Taq polymerase is
very sensitive to pH, salt concentration, etc so pipetting accurately
is critical.

2) Unless you know the concentration of your template DNA, make a
couple dilutions of your template. Start with a 100x dilution and see
if that works. FYI, high concentration of DNA can inhibit PCR or
cause non-specific amplification.

-cory

[Taylor R.]

Thanks for the reply…we'll discuss your suggestions and see what works!