SEA-PHAGES Logo

The official website of the HHMI Science Education Alliance-Phage Hunters Advancing Genomics and Evolutionary Science program.

Welcome to the forums at seaphages.org. Please feel free to ask any questions related to the SEA-PHAGES program. Any logged-in user may post new topics and reply to existing topics. If you'd like to see a new forum created, please contact us using our form or email us at info@seaphages.org.

All posts created by dedrick

| posted 16 Mar, 2016 13:01
We use a Mini-bead beater from Biospec Products, Inc. I'm not so sure vortexing would do the trick.
Bekah Dedrick
Posted in: Phage Discovery/IsolationRNA isolation protocol from smeg
| posted 16 Mar, 2016 11:39
Yes I did. I attached the protocol here.
Bekah Dedrick
Posted in: Phage Discovery/IsolationRNA isolation protocol from smeg
| posted 15 Mar, 2016 18:34
Hi Dan,
Yep, we use the Qiagen RNeasy kit. It's super easy and relatively inexpensive.
The kit has you resuspend your cell pellet in a buffer called RLT. We place this suspension in a matrix lysing tube (matrix B - MP Biomedicals 6911-100) and bead beat for 45 seconds on max speed, two times (icing in between) to break open the cells. Then, you centrifuge this tube briefly and transfer the supernatent into a new tube and go on with the procedure where you add 560ul of 80% EtOH.
Let me know if you need more information!!
Bekah
Bekah Dedrick
Posted in: Phage Discovery/IsolationRNA isolation protocol from smeg
| posted 17 Sep, 2015 00:24
Hi Lee,
Do you use this method for all Streptomyces species? Are you using liquid media specific for sporulation? I know some do not sporulate in liquid (coelicolor)…not sure if it matters. I have always used spore stocks and top agar. Of course your method sounds easier!!!!
Best,
Bekah
Bekah Dedrick
Posted in: StreptomycesStrep problems
| posted 16 Sep, 2015 14:36
Hi Steve,

Why aren't you using top agar? I have used it when isolating S. lividans phage.

Bekah
Bekah Dedrick
Posted in: StreptomycesStrep problems