Difference between revisions of "Part:BBa K1439002:Experience"
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how you used this part and how it worked out. | how you used this part and how it worked out. | ||
| − | === | + | ===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 OriTR (BBa_J01003). We got a special strain from other lab which is sensitive to Streptomycin, while Top10 has streptomycin resistance. In order to test BBa_J01003 better, we ligate OriTRP4 with the report gene RFP. After conjugation, we pick the red colonies by LB medium with resistance. | ||
| + | |||
| + | Conjugation between E.coli HB101 and Top10 | ||
| + | {| class="wikitable" | ||
| + | |- | ||
| + | ! !! Conjugation (HB101 and Top10 mixed system) !! Top10(no plasmid) !! HB101(double plasmids system) | ||
| + | |- | ||
| + | | Streptomycin || White colony || White colony || No colony | ||
| + | |- | ||
| + | | chloramphenicol || Red colony || No colony || Red colony | ||
| + | |- | ||
| + | | chloramphenicol & streptomycin || Red colony || No colony || No colony | ||
| + | |} | ||
| + | Table.1 | ||
| + | We designed a orthogonal experiment, it is showed in the table. The first column shows the strains used in coating, and the first row shows the antibiotics added in medium. Besides, the table shows the result ideally. The second row proves that HB101, plasmid RP4 and the backbone pSB1C3 don’t contain streptomycin resistant gene. The third row proves that Top10 is sensitive to chloramphenicol. And the forth row proves that Top10 can receive the mini plasmid after conjugation. | ||
===User Reviews=== | ===User Reviews=== | ||
Revision as of 02:57, 17 October 2014
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how you used this part and how it worked out.
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 OriTR (BBa_J01003). We got a special strain from other lab which is sensitive to Streptomycin, while Top10 has streptomycin resistance. In order to test BBa_J01003 better, we ligate OriTRP4 with the report gene RFP. After conjugation, we pick the red colonies by LB medium with resistance.
Conjugation between E.coli HB101 and Top10
| Conjugation (HB101 and Top10 mixed system) | Top10(no plasmid) | HB101(double plasmids system) | |
|---|---|---|---|
| Streptomycin | White colony | White colony | No colony |
| chloramphenicol | Red colony | No colony | Red colony |
| chloramphenicol & streptomycin | Red colony | No colony | No colony |
Table.1 We designed a orthogonal experiment, it is showed in the table. The first column shows the strains used in coating, and the first row shows the antibiotics added in medium. Besides, the table shows the result ideally. The second row proves that HB101, plasmid RP4 and the backbone pSB1C3 don’t contain streptomycin resistant gene. The third row proves that Top10 is sensitive to chloramphenicol. And the forth row proves that Top10 can receive the mini plasmid after conjugation.
User Reviews
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