Hello all,
I'm trying to detect the level of expression for two different mRNA with real-time PCR (SYBR green) during apoptosis. I'm using GAPDH and b-actin as my control.
But I need to change both my housekeeping genes primers as they start to show downregulation during apoptosis.
I'm looking for a substitute for these two genes. Do you think 18S would stay constant during apoptosis?
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5 replies to this topic
#2
bob1
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Posted 02 June 2009 - 04:32 PM
Ideally you will screen a few (5 or more) housekeeping genes and then choose the ones that are most stable. You could try B2M or HPRT.
#3
Rsm
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Posted 02 June 2009 - 05:36 PM
PMID 14748434 (search this number in PubMed) found that GAPDH was the best housekeeping gene during apoptosis. Also B2M was promising, while 18S was not... I don't know, maybe your cells are different from theirs? But I agree with bob1, best way is to check yourself.
Cheers,
Minna
Cheers,
Minna
I got soul, but I'm not a soldier
#4
Curtis
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Posted 02 June 2009 - 08:10 PM
ok, thank you guys.
so I'm gonna design new primers for 18s, B2M and HPRT
so I'm gonna design new primers for 18s, B2M and HPRT
#5
Rsm
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Posted 04 June 2009 - 01:26 AM
make sure to check ribosomal proteins as well... my favorite is Rpl13! Best ever, trust me. I've tried a lot.
Cheers,
Minna
Cheers,
Minna
I got soul, but I'm not a soldier
#6
Star*
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Posted 16 June 2009 - 06:05 AM
Hello! I'm facing the same problem.. both GAPDH and b-actin are affected during apoptosis... so, I would like to ask which House keeping primer worked finally ?? Please help me ...
