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Quenching the crosslinking

glycine

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#1 samit

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Posted 24 February 2012 - 09:47 AM

Hi all,

Basically I am not very new to ChIP experiment. I have been doing it very successfully since last one year but today I did make a mistake. I crosslink my cells for 10 min in 1% formaldehyde @ RT and then add gycine to stop the crosslinking and unfortunately I forgot to add glycine. and have washed them 2 times in PBS and then pelleted them in PBS+PIC, freezed them liquid nitrogen. I have to do so for several time points. after making mistake for 2 points suddenly it came to my mind , and I started to add glycine. Now I feel that I have two batches, one treated with glycien and other without it.
Please let me know if I can treat my untreated cells with same glycine amount after thawing them again without any harm??? or what could I do else in order to homogenize both batchs.

Thanks

#2 KPDE

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Posted 24 February 2012 - 10:47 PM

Just continue to process the tissues as if you had quenched them. After washing with PBS, I doubt that quenching will do very much.

#3 samit

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Posted 25 February 2012 - 01:31 AM

Thanks KPDE for your reply,

I did the same, I started the cell lysis today morning, If I will see some strange results then only I will repeat the experiment..





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