IGEM:IMPERIAL/2006/LabCalendar/2006-8-29: Difference between revisions
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<font color="#FF8000"> | <u>'''TO DO:'''</u> | ||
*Culture up the ligated parts (J37022 & J37025) for miniprep (to check whether ligations from Monday | *Culture up the ligated parts (J37022 & J37025) for miniprep (to check whether ligations from Monday | ||
were successful) from plates in incubator | were successful) from plates in incubator | ||
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More LB Amp plates ? (How many more do we need for our remaining time - there are about 10 left) | More LB Amp plates ? (How many more do we need for our remaining time - there are about 10 left) | ||
*Gels from the weekend need to be analysed: Were J37018 & J37024 ligations successful ? | *Gels from the weekend need to be analysed: Were J37018 & J37024 ligations successful ? | ||
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==LoxP PCR== | ==LoxP PCR== | ||
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*Today's gel was looked at immediately after being run and two distinct bands could be seen - however these disappeared once left for a few minutes. | *Today's gel was looked at immediately after being run and two distinct bands could be seen - however these disappeared once left for a few minutes. | ||
*Once immersed in Ethidium bromide solution, the bands reappeared under the UV light. | *Once immersed in Ethidium bromide solution, the bands reappeared under the UV light. | ||
==Ligations== | |||
*Repeat ligation J37023(Immunotagged AiiA)->1I | |||
**Digest DNA from Maxi prep stock | |||
**Run Maxi prep gel | |||
**[[Image:Maxi_j37024.pdf]] | |||
**Glass Milk Purify insert and vector (J37023 and 1I respectively) | |||
**Leave to ligate overnight | |||
==Testing== | |||
Culture Up bacteria for testing tomorrow. | |||
Latest revision as of 08:48, 30 August 2006
TO DO:
*Culture up the ligated parts (J37022 & J37025) for miniprep (to check whether ligations from Monday
were successful) from plates in incubator
*Make frozen stock of 4G(S01656), J37018 and colony 2 of J37020
*Make more 50 times TAE
*Culture for testing: T9002, J37015, J37020 (colony 2)
*To order: LB medium (or make up ourselves & send to autoclave) & make some LB Amp to have in stock
Autoclaved flasks (and cuvettes?)
Need more eppendorfs
More LB Amp plates ? (How many more do we need for our remaining time - there are about 10 left)
*Gels from the weekend need to be analysed: Were J37018 & J37024 ligations successful ?
LoxP PCR
- Run once more, this time with DMSO, and a larger amount of maxiprepped DNA
- Major breakthrough - discovered that the reason for the repeated failures maybe that the DNA is there, but is not showing up.
- Today's gel was looked at immediately after being run and two distinct bands could be seen - however these disappeared once left for a few minutes.
- Once immersed in Ethidium bromide solution, the bands reappeared under the UV light.
Ligations
- Repeat ligation J37023(Immunotagged AiiA)->1I
- Digest DNA from Maxi prep stock
- Run Maxi prep gel
- File:Maxi j37024.pdf
- Glass Milk Purify insert and vector (J37023 and 1I respectively)
- Leave to ligate overnight
Testing
Culture Up bacteria for testing tomorrow.
