I have received transformed DH5 alpha competent cell from an USA supplier that contain my target DNA. I kept it in 4 refrigerator. Now I want to transfer the culture to another lab which is 1 hour from my lab. Can I use normal ice or normal temperature should be ok keeping the cells alive and perfect for DNA isolation?
The whole point of using stab cultures is that they are stable for long periods, since the E. coli remain viable, but oxygen availability is low, thereby slowing their growth rate. Stabs are stable at RT for several weeks, and good at 4C for months. I would not worry about temperature control during transport.
Normal ice should be ok. Maybe you should avoid direct contact of the cell with the ice (e.g. like putting your culture in a second vessel (in case of an eppi containing the culture put the eppi in a 50 ml falcon tube...).
If you want to culture your cells prior plasmid isolation, transport at ambient temperature also might be ok.
best, Guido
You may prefer cryobox full with ice and make sure any contamination while transport of culture and as you have mention destination is 1 hour, it would not be problematic.
Good Luck
You can prepare and agar stab culture and transfer it at room temperature with no problem. I always ship out E. coli agar stabs at room temperature.
Hi, if its only 1 hr you should be fine at ambient temp. The doubling time for bacteria is 20-30 min but that is in optimal conditions - they won't get going straight out of the fridge anyway. It is unlikely that you'll get any significant lysis either in such a short period. However, I would make a glycerol stock, a back-up liquid culture or streak some on a plate before you go, just in case of disasters!
The whole point of using stab cultures is that they are stable for long periods, since the E. coli remain viable, but oxygen availability is low, thereby slowing their growth rate. Stabs are stable at RT for several weeks, and good at 4C for months. I would not worry about temperature control during transport.
Check out this helpful resource with all the info you need on retrieval and storage of samples http://www.addgene.org/recipient_instructions/
hi Jonnae Kamens, I actually got this culture from Addgene this week.:)
Since the place is just 1 he away transfer at ambient temperature won't be a problem.keeping in contact with ice may give freeze-thaw effect which isn't advisable.Stab or a slant would be ok.You may keep a spare transfered culture at ur lab.Best of luck.
I keep stabs at 4C for 1 week maximum. Most public repositories that I get stabs from I understood suggest that (Addgene, BACPAC, ...). I generally streak some of the stab out on a plate with appropriate antibiotics the same or the next day I get the stab, and make glycerol stocks from single colonies. After restriction verification, I keep one correct glycerol stock and the plasmid, for future use. Transferring a stab to other places are perfectly fine at room temperature. I always get packages with stabs at regular temperature.
@ Koen: Thank you very much. to check the plasmid what do you do? restriction verification? can we do colony PCR or NANO and gel electrophoresis for plasmids? how you perform the restriction verification? how to chose the enzyme?
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