Which protocol are you using?
Im just confused as to why people buy competent cells when you can easily make them yourself. Biohackers don't have money but have time so why not get a nice plasmid producing strain and make a TSS buffer based competent cell system. Mine last for 3 weeks in -20C. Works fine. I use a line of NEB turbos I bought like three years ago. Unless you have a -80 the price of comp cells are not worth it at all. Its not magic in those cells, its cell state and buffer used...all which can be obtained by the user. Commercial comp cells are just convenience for labs that absolutely need highest efficiency...though for hacker-spaces and personal labs I see no purpose in massively competent transformations. Its one of those fetishized metrics people rave about but its intrinsically pointless for most cases. After three weeks I still get 40 colonies which is more than enough to screen a ligation or colony pcr. Pardon the rant but I really don't see a point in buying when there are so many good protocols for making them simply.
Sebastian S. CociobaCEO & FounderNew York Botanics, LLCBlog: ATinyGreenCell.com--On Saturday, March 5, 2016 at 8:44:54 AM UTC-8, William Beeson wrote:--for their ultra-high competency cells they have a FAQ (https://www.neb.com/faqs/1/01/01/can-i-store-competent- ) that says after 1 week of storage at -20C the efficiency is decreased by 250x.cells-at-20-deg-c-instead-of- 80-deg-c Nice data point! They say "cells lost 94.5% of TE after only 24 hours of storage at -20°C. Cells lost 98.9% of TE after 2 days, and 99.6% of TE after one week of storage at -20°C" - so that would be 18x after one day, 91x after 2 days, and 250x after 7 days.Anyone know whether electrocompetent cells maintain their competence any better, or whether there are any particular strains that tend to maintain competence longer when stored above -80?PatrikPS: Of course, all this also points out how important shipping conditions are likely to be for competent cells
-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diybio@googlegroups.com. To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com. For more options, visit this group at https://groups.google.com/d/forum/diybio?hl=en
Learn more at www.diybio.org
---
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To unsubscribe from this group and stop receiving emails from it, send an email to diybio+unsubscribe@googlegroups.com.
To post to this group, send email to diybio@googlegroups.com.
Visit this group at https://groups.google.com/group/diybio.
To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/1d273757-64b0-440d-922f-fd55428917fd%40googlegroups.com.
For more options, visit https://groups.google.com/d/optout.
-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diybio@googlegroups.com. To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com. For more options, visit this group at https://groups.google.com/d/forum/diybio?hl=en
Learn more at www.diybio.org
---
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To unsubscribe from this group and stop receiving emails from it, send an email to diybio+unsubscribe@googlegroups.com.
To post to this group, send email to diybio@googlegroups.com.
Visit this group at https://groups.google.com/group/diybio.
To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/0A472610-3CA9-40C4-9D65-6764B2799F51%40gmail.com.
For more options, visit https://groups.google.com/d/optout.
0 comments:
Post a Comment