Composite

Part:BBa_K1123005:Experience

Designed by: Ardjan van der Linden   Group: iGEM13_TU-Eindhoven   (2013-09-08)
Revision as of 17:57, 17 October 2018 by Zefang (Talk | contribs) (→‎Applications of BBa_K1123005)


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 This characterization is provided by TJU_China 2018 IGEM team. We firstly amplified the eGFP fragment from BBa_K1123005(iGEM13_TU-Eindhoven) by PCR.

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) 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.

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