Difference between revisions of "Part:BBa K1123005:Experience"
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===Applications of BBa_K1123005=== | ===Applications of BBa_K1123005=== | ||
A characterization and usage example of this part can be found on its main page [[Part:BBa_K1123005 | BBa_K1123005]] | A characterization and usage example of this part can be found on its main page [[Part:BBa_K1123005 | BBa_K1123005]] | ||
| + | We firstly amplified the eGFP fragment from BBa_K1123005(iGEM13_TU-Eindhoven) by PCR. | ||
| + | https://static.igem.org/mediawiki/2018/e/e5/T--TJU_China--3-9.png | ||
| + | Figure 1. Amplification of eGFP fragment (BBa_K1123005) by PCR. Lane M, marker. Lane 1, 25ng eGFP fragment. Lane 2, 50ng. Lane 3, 100ng. Lane 4, 150ng. Lane 5, 200ng. | ||
| + | |||
| + | Since this fragment contains the cleavage site of our sgRNA, we used it to test the in vitro cleavage activity of our CRISPR/Cas9 system. Please notice that our cleavage site was not so suitable for this fragment since it was designed for plasmid cutting(fragment of 720bp is cut into fragments of about 80bp and 640bp) | ||
| + | https://static.igem.org/mediawiki/2018/c/ce/T--TJU_China--3-10.png | ||
| + | Figure 2. In vitro cleavage of eGFP fragment. Lane M, marker. Lane 1, eGFP fragment. Lane 2-3, sgRNA:Cas9:DNA=10:10:1. Lane 4-5, sgRNA:Cas9:DNA=10:20:1. Lane 6-7, sgRNA:Cas9:DNA=20:20:1. | ||
===User Reviews=== | ===User Reviews=== | ||
Revision as of 17:47, 17 October 2018
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_K1123005
A characterization and usage example of this part can be found on its main page BBa_K1123005
We firstly amplified the eGFP fragment from BBa_K1123005(iGEM13_TU-Eindhoven) by PCR.
Figure 1. Amplification of eGFP fragment (BBa_K1123005) by PCR. Lane M, marker. Lane 1, 25ng eGFP fragment. Lane 2, 50ng. Lane 3, 100ng. Lane 4, 150ng. Lane 5, 200ng.
Since this fragment contains the cleavage site of our sgRNA, we used it to test the in vitro cleavage activity of our CRISPR/Cas9 system. Please notice that our cleavage site was not so suitable for this fragment since it was designed for plasmid cutting(fragment of 720bp is cut into fragments of about 80bp and 640bp)
Figure 2. In vitro cleavage of eGFP fragment. Lane M, marker. Lane 1, eGFP fragment. Lane 2-3, sgRNA:Cas9:DNA=10:10:1. Lane 4-5, sgRNA:Cas9:DNA=10:20:1. Lane 6-7, sgRNA:Cas9:DNA=20:20:1.
User Reviews
UNIQ04b4f7faae04b90c-partinfo-00000000-QINU UNIQ04b4f7faae04b90c-partinfo-00000001-QINU