thanks for your feed-back,
On Thursday, 22 December 2016 20:39:07 UTC+1, Sebastian wrote:
-- i'm definitely not an expert on growing bacterial colonies, at least i'm trying to take a shower before they become to obvious LOL
i'm attaching a better quality picture. If you zoom in, you will see round dots on the CRISPR side, dots you do not see at the bottom. These are the kind of dots i had on the first plate i used to grow my normal strain (on a gel without strep). For me, these whitish streaks do not seem to be growing colonies, the streak appeared rather rapidly after i streaked both strains, once they were dried, but then they stayed like this. After 15h, i started to see the little dots on one side.
Don't know if this helps.
On Thursday, 22 December 2016 20:39:07 UTC+1, Sebastian wrote:
Picture shows both grew on LB Strep, are you sure your wild type is not resistant? The Wild Type looks far more resistant...or cell density at streaking point was higher. If you have access you may want to PCR to confirm. Also use a high fidelity polymerase since you are looking for a single nucleotide change. Either way glad you conducted the experiment. I would suggest doing a repeat or two just to be sure :)
Sebastian S. CociobaCEO & FounderNew York Botanics, LLCBlog: ATinyGreenCell.comHi guys,--i think i have successfully CRISPRed an E. Coli bacteria,I have used the kit sold here:I followed the protocol here:The gene that was targeted is Escherichia coli 30S ribosomal subunit protein S12 (rpsL) gene. One base pair was changed to render the bacteria resistant to streptomycin.The picture attached shows my result. That plate contains a gel with streptomycin. The top part shows growth on the CRISPRed strain, the bottom part shows not growth on the normal strain.What you guys think ?I'm thinking of having the DNA of that gene sequenced, just to check if the edit really happened only where expected.
-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diy...@googlegroups.com. To unsubscribe from this group, send email to diybio+un...@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+un...@googlegroups.com .
To post to this group, send email to diy...@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/f6d2b8c8-6b2f- .4dc8-90ca-f533cabb3013% 40googlegroups.com
For more options, visit https://groups.google.com/d/optout .
<IMG_20161218_091257.jpg>
-- 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/f27ca256-64dc-4967-900a-088006212e84%40googlegroups.com.
For more options, visit https://groups.google.com/d/optout.
0 comments:
Post a Comment