Difference between revisions of "Part:BBa K2515002:Experience"
| Line 2: | Line 2: | ||
__NOTOC__ | __NOTOC__ | ||
This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | ||
| − | how you used this part and how it worked out. | + | how you used this part and how it worked out.<br /><br /> |
===Applications of BBa_K2515002=== | ===Applications of BBa_K2515002=== | ||
| − | + | <p align="justify"><br /><br /> | |
| + | This part was used by our team to clone all gRNAs needed for our ACE OF BASE project. We also suggest to use the following quick and easy way to test your cloned gRNAs for functional activity. Transform this vector with your gRNA of interest into cells with Cas9 and into cells without Cas9. Having in mind that the combination of gRNA + Cas9 is lethal in <i>E. coli</i> (unless you provide repair matrix too), you should get a lot of colonies on the plate without Cas9 and none on the plate with Cas9 if everything is fine (see the picture). To use this trick your gRNA should tagret a sequence inside the <i>E. coli</i> genome.</p> | ||
| + | <br /><br /> | ||
| + | https://i.imgur.com/VopDgWt.jpg | ||
| + | <br /><br /> | ||
===User Reviews=== | ===User Reviews=== | ||
<!-- DON'T DELETE --><partinfo>BBa_K2515002 StartReviews</partinfo> | <!-- DON'T DELETE --><partinfo>BBa_K2515002 StartReviews</partinfo> | ||
Latest revision as of 20:13, 1 November 2017
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K2515002
This part was used by our team to clone all gRNAs needed for our ACE OF BASE project. We also suggest to use the following quick and easy way to test your cloned gRNAs for functional activity. Transform this vector with your gRNA of interest into cells with Cas9 and into cells without Cas9. Having in mind that the combination of gRNA + Cas9 is lethal in E. coli (unless you provide repair matrix too), you should get a lot of colonies on the plate without Cas9 and none on the plate with Cas9 if everything is fine (see the picture). To use this trick your gRNA should tagret a sequence inside the E. coli genome.
User Reviews
UNIQ50dcb5e5b694b6e3-partinfo-00000000-QINU UNIQ50dcb5e5b694b6e3-partinfo-00000001-QINU