Difference between revisions of "Part:BBa K1918104:Experience"

Latest revision as of 21:17, 19 October 2016

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_K1918104

Firstly, we need to acquire the elements. We using PCR, Overlap PCR and Gibson to get the elements such as EBFP2, EBFP2N(1-154), EBFP2C(155-238), EBFP2N(1-154):IntN and IntC:EBFP2(155-238).

Secondly, we construct the part. We combine the elements with the vector and transfer the vector into the competent E. coli cells. After that, we select the positive clones and use colony PCR to identify the recombinants. Culturing the positive clones and sequencing the isolated plasmids to make sure that we have get the needed parts.

Lastly, we transfect the plasmids to 289H cells and do the FACS test. In the results of FASC test, the EBFP-A signals tell us that the split florescent proteins could fuse together and our parts work fine.

User Reviews

UNIQ3f5bac13358d58a5-partinfo-00000000-QINU UNIQ3f5bac13358d58a5-partinfo-00000001-QINU

@@ Line 5: / Line 5: @@
 ===Applications of BBa_K1918104===
+Firstly, we need to acquire the elements.
+We using PCR, Overlap PCR and Gibson to get the elements such as EBFP2, EBFP2N(1-154), EBFP2C(155-238), EBFP2N(1-154):IntN and IntC:EBFP2(155-238).
+Secondly, we construct the part.
+We combine the elements with the vector and transfer the vector into the competent E. coli cells. After that, we select the positive clones and use colony PCR to identify the recombinants. Culturing the positive clones and sequencing the isolated plasmids to make sure that we have get the needed parts.
+Lastly, we transfect the plasmids to 289H cells and do the FACS test.
+In the results of FASC test, the EBFP-A signals tell us that the split florescent proteins could fuse together and our parts work fine.
 ===User Reviews===