From 2013.igem.org

Revision as of 11:23, 11 July 2013

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

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.

After purification of AMO, HAO, CYC the results can be shown on the gel below.