Team:Marburg/Notebook:April

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Notebook: April

26.04.2013

Sequencing
Investigator: Franzi
Aim: Examination of sequencing results (23.4.13).

1C3 iBB1 K1 + K2 correct

1C3 iBB4 K1 + K2 correct

1C3 iBB7 K3 + K4 correct

1C3 iBB5 K1 + K2 Mutagenesis of XhoI-Site failed, only necessary for iGEM-standard 10 → unnecessary, rest correct

1C3 iBB3 K1 + K2 same point-Mutation → template?

1C3 iBB6 K1 + K2 same point-mutation → template?

Sequencing
Investigator: Franzi
Aim: Sequencing of pPhaNR.

Original pPhaNR was sent to sequencing using primers imR25/26 for iBB3 and iBB6.

Miniprep
Investigator: Franzi
Aim: TBA

PSB1C3 iBB8 K1 and pSB1C3 iBB2 K1-4 were isolated via Miniprep, eluted in 40 l Elutionbuffer.

Digest
Investigator: Franzi
Aim: TBA
  • 1 µl DNA template
  • 0,5 µl EcoRI
  • 0,5 µl Pst
  • 1 µl Cut Smart buffer
  • 6x loading buffer
  • 7 µl H2O

The samples were incubated for 1 h at 37° C.

Gel electrophoresis
gel-electrophoresis-image

Gel substances

  • 1% Agarose gel
  • 10 µl RedSafe per 50 ml gel
  • 6 µl GeneRuler™ 1kb plus DNA Ladder (Thermo Scientific)

IBB8 negative, iBB2 all positive.
Sequencing
Investigator: Franzi
Aim: Sequencing of pSB1C3 iBB2.

IBB2 K2 + K4 sequenced with primer VF2_fwd and VR_rev.

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