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
0
Lysis method for single human embryonic cell
Started by badguy, Sep 23 2012 11:33 PM
embryonic cell lysis buffer
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 advance
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
#2
bob1
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Thelymitra pulchella
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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?
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. lol
I'l update you guys after do the PCR
I'l update you guys after do the PCR
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