"
Team:DTU-Denmark-2/Notebook/Lab notes
From 2011.igem.org
(→August) |
|||
| Line 81: | Line 81: | ||
The U2OS cells are splitted and passed on to two new 75 cm2 culture flasks. | The U2OS cells are splitted and passed on to two new 75 cm2 culture flasks. | ||
Hopefully, they will be ready to pass on to cover slips on monday. | Hopefully, they will be ready to pass on to cover slips on monday. | ||
| + | |||
| + | ==='''Monday 15.08.2011'''=== | ||
| + | The U2OS are splitted and passed on to coverslips placed in a 6-well plate and a new 75 cm2 flask. They are placed in the incubator overnight - and tomorrow they will be ready for transfection. | ||
| + | |||
| + | ==='''Tuesday 16.08.2011'''=== | ||
| + | Transfection of U2OS cells. | ||
=='''September'''== | =='''September'''== | ||
Revision as of 08:34, 16 August 2011
Contents |
Lab notes
June
July
Tuesday 19.07.2011
First day in the lab and 48 biobricks to go, how exciting!
Of the 48 biobricks we will have when the project is finished we had only established 17 of them and ordered primers. We started the day with 11 PCR reactions of the first bash of 17 biobricks, we still need some DNA templates to due all 17. 6 PCR were correct and was purified with a purification Kit (GE Healthcare). So only 42 biobricks to go!
Biobricks in the bag:
- pALC
- TtrpC
- Terminator 1
- Terminator 2
- Terminator 3
- argB
- Insert GEL Photo
Thursday 21.07.2011
Some of the PCR reactions that didn't work Tuesday was repeated with a different annealing temperature and amplification of new biobricks.
New biobricks in the bag:
- Insert of gel photo
- PGK
- BGHpA
- eGFP+k_GOI
The fragments will be frozen and purified later.
We still have not succeed to make biobricks pyrG and pyrG-DR (use as a marker in fungi), despite two PCR attempts with a high annealing (59°) and low (56°) temperature.
Work for iGEM Copenhagen team
We have established a collaboration with iGEM Copenhagen team, where we will create some biobricks for them and they can use our plug 'n' play system and they will save a lot of time.
So today we transformed competent E.coli cells with some of the standard biological parts from iGEM, so we later take make biobricks to match our system.
Friday 22.07.2011
Monday 25.07.2011
Tuesday 26.07.2011
Wednesday 27.07.2011
August
Thursday 04.08.2011
Today we purified 22 biobricks, which have been obtained over the last week of laboratory work with PCR. The purification of DNA was done by using the illustra GFX PCR DNA and Gel Band Purification Kit (GE Healthcare) according to manufactures manual for purification of DNA from TAE and TBE agarose gels protocol, see protocol section for more information.
Today, we also started cultivation of the two mammalian cell lines, HeLa and U2OS. They were defrosted and cultivated in in DMEM medium containing serum, penicillin and streptomycin. Each cell line was then transferred to a 25 cm2 culture flask, see protocol section.
- pyrG TEMP
Friday 05.08.2011
Monday 08.08.2011
U2OS cells HeLa cells
Wednesday 10.08.2011
U2OS cells HeLa cells
Friday 12.08.2011
The U2OS cells are splitted and passed on to two new 75 cm2 culture flasks. Hopefully, they will be ready to pass on to cover slips on monday.
Monday 15.08.2011
The U2OS are splitted and passed on to coverslips placed in a 6-well plate and a new 75 cm2 flask. They are placed in the incubator overnight - and tomorrow they will be ready for transfection.
Tuesday 16.08.2011
Transfection of U2OS cells.
September
