we already tried 18S and beta-actin in our lymphnode samples - but they are highly regulated, which makes our calculations quite difficult.
anyone of you experience with other housekeeping genes??
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#2
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Posted 03 November 2009 - 09:31 AM
moljul, on Nov 3 2009, 11:04 AM, said:
we already tried 18S and beta-actin in our lymphnode samples - but they are highly regulated, which makes our calculations quite difficult.
anyone of you experience with other housekeeping genes??
anyone of you experience with other housekeeping genes??
GAPDH, a- or B- tubulin, and HPRT come to mind as options. I'd recommend checking the literature for other people who have done molecular work on the same treatment, and see what they use as a HKG. If there's not much out there, you may have to try a bunch of genes and see what has the least variation. There are a couple programs that will find the lowest variance among multiple genes and samples.
Above all things, if kindness is your king,
then heaven will be yours, before you meet your end
then heaven will be yours, before you meet your end
#3
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Posted 03 November 2009 - 11:07 AM
gfischer, on Nov 3 2009, 09:31 AM, said:
moljul, on Nov 3 2009, 11:04 AM, said:
we already tried 18S and beta-actin in our lymphnode samples - but they are highly regulated, which makes our calculations quite difficult.
anyone of you experience with other housekeeping genes??
anyone of you experience with other housekeeping genes??
GAPDH, a- or B- tubulin, and HPRT come to mind as options. I'd recommend checking the literature for other people who have done molecular work on the same treatment, and see what they use as a HKG. If there's not much out there, you may have to try a bunch of genes and see what has the least variation. There are a couple programs that will find the lowest variance among multiple genes and samples.
thanks, gfischer!!
#4
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Posted 04 November 2009 - 03:27 AM
there are companies that are selling HKG panels for testing on your respective samples like here or here.
You can also try to do this with your own primers. There are a lot of published HKG primer sequences available like here.
Good luck!
You can also try to do this with your own primers. There are a lot of published HKG primer sequences available like here.
Good luck!
tea-test: The artist formerly known as Ned Land
#5
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Posted 09 November 2009 - 10:16 AM
Hi,
I'm also searching for housekeeping genes, I have heard bad things about b-actin and GAPDH (That they vary quite alot..)so was going to try RP11 and G6PD or Rpia, has anyone had any trouble with these, I cant find much literature on qPCR for my treatment...
Thanks for any ideas or probs encountered.
SciS
I'm also searching for housekeeping genes, I have heard bad things about b-actin and GAPDH (That they vary quite alot..)so was going to try RP11 and G6PD or Rpia, has anyone had any trouble with these, I cant find much literature on qPCR for my treatment...
Thanks for any ideas or probs encountered.
SciS
