Difference between revisions of "Part:BBa K1439000:Experience"
(→Test1. Conjugation between E.coli HB101 and Top10) |
(→Test1. Conjugation between E.coli HB101 and Top10) |
||
Line 1: | Line 1: | ||
__NOTOC__ | __NOTOC__ | ||
===Test1. Conjugation between E.coli HB101 and Top10=== | ===Test1. Conjugation between E.coli HB101 and Top10=== | ||
− | + | We do conjugation experiment between HB101 and Top10 to test the conjugation ability of OriTRP4 (BBa_K1439000). We got a special strain from other lab which is sensitive to Streptomycin, while Top10 has streptomycin resistance. In order to test BBa_K1439000 better, we ligate OriTRP4 with the report gene RFP. After conjugation, we pick the red colonies by LB medium with resistance. | |
− | + | ||
− | + | ||
===Applications of BBa_K1439000=== | ===Applications of BBa_K1439000=== |
Revision as of 12:30, 16 October 2014
Test1. Conjugation between E.coli HB101 and Top10
We do conjugation experiment between HB101 and Top10 to test the conjugation ability of OriTRP4 (BBa_K1439000). We got a special strain from other lab which is sensitive to Streptomycin, while Top10 has streptomycin resistance. In order to test BBa_K1439000 better, we ligate OriTRP4 with the report gene RFP. After conjugation, we pick the red colonies by LB medium with resistance.
Applications of BBa_K1439000
We have test the oriTRP4 BBa_K1439000 in a conjugation system in which pSB1C3 with the help of plasmid RP4, and E. coli HB101 as donor and E.coil Top10 as recipient cells. We test the conjugative competence of oriTRP4 BBa_K1439000 by conjugate between HB101 and Top10. We use the RFP (BBa_J04450) marks the plasmids containing oriTRP4 BBa_K1439000. We get HB101 without streptomycin resistance form Zhigang Qiu. Top 10 has streptomycin resistance. After conjugation, we select recipient cells by streptomycin resistance LB medium and statistical number of red bacterial colony.
User Reviews
UNIQ791eb1a2c2f5ebcf-partinfo-00000000-QINU UNIQ791eb1a2c2f5ebcf-partinfo-00000001-QINU Test
Conjugate between E.coil HB101 and E.coil Top10.