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From 2013.igem.org

Contents 1 208 1.1 Main purpose 1.2 Who was in the lab 1.3 Procedure 1.3.1 PCR in order to amplify AMO and HAO with USER primers 1.3.2 gel electrophoresis 1.3.3 ON cultures and re-plating of Nir USER transformants from Colony PCR 1.4 Results 1.4.1 Gel 1 1.4.2 Gel 2 1.5 Conclusion

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Main purpose

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• 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

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Kristian, Gosia, Julia

Procedure

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PCR in order to amplify AMO and HAO with USER primers

gel electrophoresis

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

Positive Nir USER transformants obtained by colony PCR:

• Samples 25 and 26 were inoculated in 5 mL LB medium + 30 ug/ml kanamicyn for ON cultures preparation.
• Samples 25, 26 and 27 were re-plated in LB agar + 30 ug/ml kanamicyn

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

Gel 2

Conclusion

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