Difference between revisions of "Part:BBa K4630114:Experience"
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| − | Also, we performed time-gradient tests over the part | + | Also, according to our PI's advice, IPTG induction do harm to the cells. However, refractory phase is a key parameter of the multi-level recorder. So, we performed time-gradient tests over the part (tbl 1). |
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| − | <div class="col-lg" style="margin:auto;text-align: | + | All of the samples achieve full knock-out, at a rate of 100% (fig 2). However, the time-gradient within 2 hours did not really set up, because of inconsistency with the time-gradient tes in BBa_K4630100. Not surprisingly, the test carried out on stgRNA-cassette (1+2+3) showed some kind of leakage expression. More experiments of different promoters should be taken. |
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| − | + | Table 1 Time-gradient test | |
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| + | <img style="margin:5px auto 5px auto;" src="https://static.igem.wiki/teams/4630/wiki/parts/bba-k4630100-stgrna-barcode-cassette-1/table10.png" width="60%"> | ||
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Latest revision as of 23:56, 11 October 2023
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_K4630114
This part is the more integrated form that contains three cassettes. Using this part, we can verify the concept of multi-level recording and gain comprehensive data.
General Induction Protocol
- Pick a colony from the verified plate, cultivate in 5ml liquid LB media for 12 hours.
- Transfer 100µl bacteria solution to 3ml new liquid LB media. Cultivate to an OD600 of 0.3~0.5.
- Add L-Arabinose and IPTG to final concentrations of 2g/L and 3g/L, and induce for 22 hours and 5 hours, respectively.
- Spread the diluted solution to plates with appropriate antibiotics and cultivate for 12 hours.
- Pick the colonies and test the editing result.
Result
Proof-of-Concept
We implement induction over the part, and it is successfully knock-out.
Fig 1 The successful knock-out of the stgRNA-cassette (1+2+3)
Time-Gradient Test
Also, according to our PI's advice, IPTG induction do harm to the cells. However, refractory phase is a key parameter of the multi-level recorder. So, we performed time-gradient tests over the part (tbl 1).
All of the samples achieve full knock-out, at a rate of 100% (fig 2). However, the time-gradient within 2 hours did not really set up, because of inconsistency with the time-gradient tes in BBa_K4630100. Not surprisingly, the test carried out on stgRNA-cassette (1+2+3) showed some kind of leakage expression. More experiments of different promoters should be taken.
Table 1 Time-gradient test
Fig 2 Time-gradient test over the part
User Reviews
UNIQc26649004b6bfba6-partinfo-00000003-QINU UNIQc26649004b6bfba6-partinfo-00000004-QINU