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Lysis method for single human embryonic cell

embryonic cell lysis buffer

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5 replies to this topic

#1 badguy

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Posted 23 September 2012 - 11:33 PM

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 advancePosted Image

#2 bob1

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Posted 24 September 2012 - 01:16 AM

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.

#3 Curtis

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Posted 24 September 2012 - 01:47 AM

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.

#4 badguy

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Posted 24 September 2012 - 02:01 AM

I dont know any DNase inhibitor that would not interfere PCR.
any advice?

#5 bob1

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Posted 24 September 2012 - 12:52 PM

Good point - don't worry about DNase inhibitors then, they aren't needed for colony PCR, so probably not here either.

#6 badguy

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Posted 24 September 2012 - 06:58 PM

I never thought of that. lolPosted Image
I'l update you guys after do the PCRPosted Image





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