Posted 14 April 2004 - 06:06 PM
I dont know what happened with SSE induction. anyone would like give some suggestions about it.
Posted 14 April 2004 - 07:04 PM
Posted 14 April 2004 - 09:01 PM
In order to get RNA as much as possible, the bacterias were grown in the 200ml of YMB medium, so i must collect it by centrifugation. I will try to do it with higher rpm in a shorter time period.
Posted 15 April 2004 - 05:05 AM
Posted 15 April 2004 - 05:02 PM
I isolated the RNA yesterday and found that unbelievable yields of pellets came out after precipitation with isopropanol ,but it was not completely resuspended in RNase-free water.
The gel-running clearly showed the band of 23S and 16S, but the band before 16S made me disppointed . I was not sure if the mRNA had already degraded , so i want to ask how to know it ?
Posted 15 April 2004 - 07:05 PM
Go ahead and proceed with your experiment. Don't worry about what you see (or don't see) on the gel. It is a bacterial population, your cells are in mixed growth phase (some are in early log phase, some are in late stat phase, some are in death phase), the nett mRNA content per cell is going to be highly variable, so yields are going to be low. However your photo or radio probes will work easily enough- start with a dot blot.
Good luck and God Bless