"
Team:DTU-Denmark/Notebook/8 July 2013
From 2013.igem.org
Contents |
208
Main purpose
Run gel with 1 uL AMO, 10 uL AMO, 20 uL AMO genes from "Nitrosomonas Europaea". Purification and extraction of the gel for 10 uL AMO. Run 2nd gel with all the sample of 10 uL AMO. Run gel with HAO and CYC genes "Nitrosomonas Europaea".
Who was in the lab
Kristian, Ariadni, Gosia
Procedure
Run 1st gel (AMO):
- 1ul of dye
- 5ul of sample
- 5ul Kb ladder
Purification of first gel for 10 AMO according to the protocol QIA gel extraction protocol. (given in the Kit)
Run 2nd gel (AMO):
- 9ul of dye
- around 20 ul of each sample
- 12 ul of ladder
Purification of second gel according to the same protocol.
Measure the concentration of the purified AMO using Nanodrop (pouring together both samples).
Run 3rd gel (HAO,CYC):
- 1ul of dye
- 5ul of sample
- 5ul Kb ladder
Purification of DNA of both HAO and CYC.
Results
Nanodrop measurement:
- 30.4 ng/ul (AMO)
- 14.4 ng/ul (HAO)
- 11.7 ng/ul (CYC)
Pictures from gels
Gel AMO
Wells:
- 1: 1 AMO
- 2: 1 AMO
- 3: 10 AMO
- 4: 10 AMO
- 5: 20 AMO
- 6: 20 AMO
- 7: 1 Kb Plus DNA Ladder
Conclusion for AMO
Tha bands for 10 AMO are thick and it is not visible how many base-pairs they have, so gel extraction and purification is needed.
Gel HAO-CYC <hv> Wells:
- 1: 1 Kb Plus DNA Ladder
- 2: 3 HAO
- 3: 6 HAO
- 4: 10 HAO
- 5: 3 HAO
- 6: 6 HAO
- 7: 10 HAO
Around 3000 base-pairs
- 8: 3 CYC
- 9: 6 CYC
- 10: 10 CYC
- 11: 3 CYC
- 12: 6 CYC
- 13: 10 CYC
Around 1650 base-pairs
- 14: 1 Kb Plus DNA Ladder
Conclusion for HAO and CYC
The bands have shown that the results are close to 2866 bp for HAO and close to 1497 bp for CYC.
