Lysis method for single human embryonic cell - (Sep/24/2012 )
I need help from you guys regarding lysis method for single human embryonic cell.
Other than using Proteinase K and mild detergent (eg. tween-20), is it posible to use boiling lysis method or heat denaturation just like in colony PCR?
I will use this sample to do PCR. Thanks in advance
They should both work - eukaryotic cells are weak compared to prokaryotic so I don't see why not. You might need to add some DNase inhibitor or something.
You don't even need to pre-boil it. Just directly add it into your PCR reaction. It will break in the first denaturation step.
I dont know any DNase inhibitor that would not interfere PCR.
any advice?
Good point - don't worry about DNase inhibitors then, they aren't needed for colony PCR, so probably not here either.
I never thought of that. lol
I'l update you guys after do the PCR