Difference between revisions of "Part:BBa K4630113:Experience"
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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 | ||
how you used this part and how it worked out. | how you used this part and how it worked out. | ||
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===Applications of BBa_K4630113=== | ===Applications of BBa_K4630113=== | ||
This part is integrated and contains two cassettes. As the intermediate of the single cassette and multiple cassettes, BBa_K4630113 functions as the trouble shooting tool and helps the primary proof-of-concept of the cascade system. | This part is integrated and contains two cassettes. As the intermediate of the single cassette and multiple cassettes, BBa_K4630113 functions as the trouble shooting tool and helps the primary proof-of-concept of the cascade system. | ||
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====Result==== | ====Result==== | ||
| − | The | + | The knock-out of the first cassettes were successful(fig 1a). However, the second cassette remained intact. In line with the previous data from stgRNA-cassette 1, we realized that there might be some technical obstacles of the knock-out of the last cassette of the device. Comparing with stgRNA-barcode-cassette 1, we have more confidence that the homologous arms play a important role. (see also documentations of BBa_K4630110[https://parts.igem.org/Part:BBa_K4630110]) . |
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Fig 1 The induction knock-out results of stgRNA-cassette (1+2) and stgRNA-cassette 1<br> | Fig 1 The induction knock-out results of stgRNA-cassette (1+2) and stgRNA-cassette 1<br> | ||
| − | (a) Induction knock-out result of stgRNA-cassette (1+2) | + | (a) Induction knock-out result of stgRNA-cassette (1+2).<br> |
| − | (b) Induction knock-out | + | (b)Induction knock-out result of stgRNA-cassette 1, All of them remained intact. |
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Latest revision as of 00:59, 12 October 2023
Applications of BBa_K4630113
This part is integrated and contains two cassettes. As the intermediate of the single cassette and multiple cassettes, BBa_K4630113 functions as the trouble shooting tool and helps the primary proof-of-concept of the cascade system.
Experiments
The experiment we carried out on BBa_K4630113 is mainly the induction and verification.
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
The knock-out of the first cassettes were successful(fig 1a). However, the second cassette remained intact. In line with the previous data from stgRNA-cassette 1, we realized that there might be some technical obstacles of the knock-out of the last cassette of the device. Comparing with stgRNA-barcode-cassette 1, we have more confidence that the homologous arms play a important role. (see also documentations of BBa_K4630110[1]) .
Fig 1 The induction knock-out results of stgRNA-cassette (1+2) and stgRNA-cassette 1
(a) Induction knock-out result of stgRNA-cassette (1+2).
(b)Induction knock-out result of stgRNA-cassette 1, All of them remained intact.
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