Team:Heidelberg/k4l1um

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6-15-2009

Lab: LV, SH, CZ

SH:

  • Miniprep of GFP template plasmid, pcDNA5/FRT
Nr pcDNA5/FRT GFP
1 4,7 39,7
2 5,9 14,9
3 3,7 3,8
4 5,0 7,8


LV:

  • Extraction of CMV promoter from 2008 distribution
  • Transformation of DH5a cell with CMV promoter
  • portzughtr

6-16-2009

Lab: LV, SH, CZ

  • Annealing of Oligos (JeT, cFos, Min)
  • Purification of cDNA via 2% / 3% agarosegel

=> Only bands at ca. 50 Bp => no successful amplification

6-17-2009

Lab: LV, SH

  • No transformations could be observed
  • Replace Phsuion stocks to Phusion Master Mix from Nathan
  • Repeat Annealing of Oligos with a different protocol: 1:10 - 1:1000 dilution of Oligos, 95° 5', 7* [95° 45 58° 45 72° 1'], add 1:10 diluted primers, 95° 5' 25* same temperatures
  • Repeat PCR of pcDNA5/FRT and mcherry with lower annealing temperature (58°C) and fresh Polymerase