I just bought the "subcloning efficiency" competent cells from NEB (https://www.neb.com/products/c2988-neb-5-alpha-competent-e-coli-subcloning-efficiency) and they don't specifically talk about competency dropping for these cells, but for their ultra-high competency cells they have a FAQ (https://www.neb.com/faqs/1/01/01/can-i-store-competent-cells-at-20-deg-c-instead-of-80-deg-c) that says after 1 week of storage at -20C the efficiency is decreased by 250x.
-- I don't need high competency. If I have >1x10^6 CFU/ug that should work, maybe even a lot lower would work. Have you ever checked to see if the efficiency of your cells were actually dropping over time? I wonder if the drops are only for ultra high competency cells (>1x10^9 cfu/ug).
On Friday, March 4, 2016 at 2:36:57 PM UTC-5, Sebastian wrote:
On Friday, March 4, 2016 at 2:36:57 PM UTC-5, Sebastian wrote:
I make em every three weeks. Store in -20C in TSS buffer. I make how much I normally use in two weeks plus minus a bit. It isn't a big deal. For super tricky ligations I make em fresh day of. Never missed a clone. How many colonies do you really need, though?
Sebastian S. CociobaCEO & FounderNew York Botanics, LLCBlog: ATinyGreenCell.comHi all,
Any ideas on how to store moderate competency cells without a -80c freezer? I've read the cfu/ug goes down dramatically if stored at only -20c? How are you guys doing it?
Thanks,
-Will
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