Difference between revisions of "Part:BBa K341458:Experience"
| Line 1: | Line 1: | ||
| − | |||
__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 | ||
| Line 5: | Line 4: | ||
===Applications of BBa_K341458=== | ===Applications of BBa_K341458=== | ||
| + | This part is a Insert Fragment of Donor Plasmid in the In-vivo Recombination System of iGEM team Tsinghua. | ||
| + | |||
| + | At the 5’ end of Chlr, I-scel recognizing sequence and recombination sequence 1 is added. At the 3’ end, we add another recombination sequence and also I-scel recognizing sequence. | ||
| + | |||
| + | After construction, we will transform this donor plasmid to E.coli with landing pad and helper plasmid. After arabinose and IPTG inducing, the restriction enzyme I-scel will cut down chlr (containing recombination sequences), which can recombine with the bacterial chromosome. | ||
| + | |||
| + | Then we using two primers to identify the recombination of this Insert Fragment. The upper primer is complement to the genome near the recombination site, the downstream primer is complement to the Chl resistance gene. | ||
| + | |||
| + | So positive result of this PCR can prove that the recombination is workable, thus this part is workable. | ||
| + | |||
| + | [[Image:Result of Part Km for Tsinghua.jpg]] | ||
===User Reviews=== | ===User Reviews=== | ||
Latest revision as of 16:22, 6 November 2010
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_K341458
This part is a Insert Fragment of Donor Plasmid in the In-vivo Recombination System of iGEM team Tsinghua.
At the 5’ end of Chlr, I-scel recognizing sequence and recombination sequence 1 is added. At the 3’ end, we add another recombination sequence and also I-scel recognizing sequence.
After construction, we will transform this donor plasmid to E.coli with landing pad and helper plasmid. After arabinose and IPTG inducing, the restriction enzyme I-scel will cut down chlr (containing recombination sequences), which can recombine with the bacterial chromosome.
Then we using two primers to identify the recombination of this Insert Fragment. The upper primer is complement to the genome near the recombination site, the downstream primer is complement to the Chl resistance gene.
So positive result of this PCR can prove that the recombination is workable, thus this part is workable.
User Reviews
UNIQ20d14f3e62c35448-partinfo-00000000-QINU UNIQ20d14f3e62c35448-partinfo-00000001-QINU