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Revision as of 12:36, 25 July 2013

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Main purpose

• PCR of AMO and HAO using USER primers and as a template using AMO and HAO extracted from chromosomal DNA with non-uracil primers
• verification of PCRs
• ON cultures and re-plating of Nir USER transformants from Colony PCR ( 17-07-2013)

Who was in the lab

Kristian, Gosia, Julia

Procedure

PCR in order to amplify AMO and HAO with USER primers

gel electrophoresis

1% agarose gel was loaded with samples as follows:

1. 1 kb ladder
2. AMO 5uL template, 1
3. AMO 5uL template, 2
4. AMO 10uL template, 1
5. AMO 10uL template, 2
6. HAO 5uL template, 1
7. HAO 5uL template, 2
8. HAO 10uL template, 1
9. HAO 10uL template, 2
10. restriction analysis cyc EcoRI
11. restriction analysis cyc HindIII
12. 1 kb ladder

ON cultures and re-plating of Nir USER transformants from Colony PCR

2 positive Nir USER transformants obtained by colony PCR were inoculated in 5 mL LB medium + 30 ug/ml kanamicyn for ON cultures preparation

Results

Gel 1

1. 1 kb ladder
2. AMO 5uL template, 1
3. AMO 5uL template, 2
4. AMO 10uL template, 1
5. AMO 10uL template, 2
6. HAO 5uL template, 1
7. HAO 5uL template, 2
8. HAO 10uL template, 1
9. HAO 10uL template, 2
10. restriction analysis cyc EcoRI
11. restriction analysis cyc HindIII
12. 1 kb ladder

600px| pic

Conclusion

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