IGEM:Peking/2007/Count-Conjugation-Notebook/2007-7-16: Difference between revisions
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(New page: ===oriT Knock Out=== *By Xu Anting and Liu Ting :Cannot repeat the PCR result of oriT in F/R751/pSC101. In the four trials with various conditions, F bands appeared twice, R751 once, and ...) |
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*By Liu Ting and Xu Anting | *By Liu Ting and Xu Anting | ||
:Cells can grow up in LB medium with 37 centigrade, overnight. Still cannot make sure whether the conjugation plasmids are in the grown cells because of the PCR failure. | :Cells can grow up in LB medium with 37 centigrade, overnight. Still cannot make sure whether the conjugation plasmids are in the grown cells because of the PCR failure. | ||
===Eletrophoresis of Extracted Plasmid=== | |||
*By Ma and Yu Tao. | |||
*J23066, pSB3K3, pSB1AK3, R0010, J61003 are all tested. (Without Digestion.) | |||
*Each of R0010, J61003 and pSB1AK3 has a single band. (Weird) And pSB3K3 and J23066 have no bands. | |||
*Notes: This eletrophoresis is especially for me to better understand the essence of electrophoresis, to make better use of the DNA dye and to adjust the apparatus to a better condition. Besides, there is still something wrong with our UVP Bioimaging System. Therefore, the result of the electrophoresis cannot be shown. | |||
===Miniprep of E0040=== | |||
*BY Yu Tao. | |||
===Eletrophoresis of Extracted Plasmid=== | |||
*By Yu Tao. | |||
*R0040, J01060, E0040 adn B0015 are tested. (Without Digestion.) | |||
*All have a single band. | |||
*Note: | |||
==Instruction of the DNA Dye== | |||
For conveniences, the DNA dye can be diluted 100 times by TAE first and stored at 4 centigrade degree. The final cencentration in the Sample can be 1000 times diluted. | |||
===Preculture E0040 J23066 and pSB3K3 for Miniprep Tomorrow=== | |||
*By Yu Tao. | |||
Revision as of 09:06, 16 July 2007
oriT Knock Out
- By Xu Anting and Liu Ting
- Cannot repeat the PCR result of oriT in F/R751/pSC101. In the four trials with various conditions, F bands appeared twice, R751 once, and pSC101 twice.
- To-do list: use chromosome extraction template, Taq and Pfu enzyme to try again. Plan to re-design primers in case it fails again.
Strain cultivation
- By Liu Ting and Xu Anting
- Cells can grow up in LB medium with 37 centigrade, overnight. Still cannot make sure whether the conjugation plasmids are in the grown cells because of the PCR failure.
Eletrophoresis of Extracted Plasmid
- By Ma and Yu Tao.
- J23066, pSB3K3, pSB1AK3, R0010, J61003 are all tested. (Without Digestion.)
- Each of R0010, J61003 and pSB1AK3 has a single band. (Weird) And pSB3K3 and J23066 have no bands.
- Notes: This eletrophoresis is especially for me to better understand the essence of electrophoresis, to make better use of the DNA dye and to adjust the apparatus to a better condition. Besides, there is still something wrong with our UVP Bioimaging System. Therefore, the result of the electrophoresis cannot be shown.
Miniprep of E0040
- BY Yu Tao.
Eletrophoresis of Extracted Plasmid
- By Yu Tao.
- R0040, J01060, E0040 adn B0015 are tested. (Without Digestion.)
- All have a single band.
- Note:
Instruction of the DNA Dye
For conveniences, the DNA dye can be diluted 100 times by TAE first and stored at 4 centigrade degree. The final cencentration in the Sample can be 1000 times diluted.
Preculture E0040 J23066 and pSB3K3 for Miniprep Tomorrow
- By Yu Tao.
