 searching housekeeping genes |
  |
 Nov 3 2009, 09:04 AM
Post
#1
|
|
|
Enthusiast  Group: Active Members Posts: 66 Joined: 27-January 09 From: Austria Member No.: 6575  |
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?? |
 |
 
|
|
Nov 3 2009, 09:31 AM
Post
#2
|
|
 Veteran Group: Active Members Posts: 116 Joined: 26-January 09 From: Milwaukee, WI Member No.: 6465 |
QUOTE (moljul @ Nov 3 2009, 11:04 AM)  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?? 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. -------------------- Be wary of those who believe in a neat little world,
'cause it's just f*#^$ing crazy, you know that it is |
|
|
|
|
Nov 3 2009, 11:07 AM
Post
#3
|
|
|
Enthusiast Group: Active Members Posts: 66 Joined: 27-January 09 From: Austria Member No.: 6575 |
QUOTE (gfischer @ Nov 3 2009, 09:31 AM) QUOTE (moljul @ Nov 3 2009, 11:04 AM) 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?? 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!! |
|
|
|
|
Nov 4 2009, 03:27 AM
Post
#4
|
|
|
Enthusiast Group: Active Members Posts: 49 Joined: 26-January 09 Member No.: 6378 |
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! -------------------- tea-test: The artist formerly known as Ned Land
|
|
|
|
|
Nov 9 2009, 10:16 AM
Post
#5
|
|
|
member Group: Active Members Posts: 16 Joined: 22-May 09 Member No.: 9716 |
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 |
|
|
|
|
|
Lo-Fi Version | Time is now: 22 Nov 2009 - 09:06 AM |
| IP.Board Skin Developed By Creative Networks | ||
|
Home
- About
- Terms of Service
- Privacy
- Feedback
©1999-2009 Protocol Online, All rights reserved. |