Team:DTU-Denmark/Notebook/25 July 2013

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==Conclusion==
==Conclusion==

Revision as of 12:41, 25 July 2013

Contents

208

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



2013-07-25 AMO HAO and restriction cyc EcoRI HindIII.jpg

Conclusion

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