Low RNA yield
tyson at canada.com
Fri Feb 4 17:21:42 EST 2000
...perhaps there is RNase contamination somewhere eg. pipette tips etc..
"Karen Roberts" <leehrat at mindspring.com> wrote in message
news:3898A662.12D138BA at mindspring.com...
| I am posting for a technician friend who doesn't have Internet access
| (one of the few).
| She says that she is getting low yields of RNA from thymus cell
| culture. She uses Qiagen RNAeasy kit. The cell number in her samples
| is 1 x 10(7), which is the max starting amount recommended.
| She has repeated the assay three times and it appears to be a systematic
| problem. She says she is careful to homogenize the cells well acc. to
| protocol; in short her methods are fine. Though she didn't run an RNA
| gel to see how abundant it is, she did do RT PCR for B Actin and got a
| signal, but it was much less intense than what is usual for Actin. So
| she does have RNA just a low amount.
| She then made fresh buffer from the kit with fresh B 2ME and still got
| the same results.
| Could it be the kit has gone bad? She received it new from Qiagen in
| September. IF not, what else could she try to increase her yields.
| PS. This assay was done by another tech shortly after the kit arrived in
| Sept. and she had the expected amt of RNA. Though that would suggest
| it's my friend's technique, she asked the investigator to repeat it and
| she also got low yields.
| Ideas appreciated.
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