Team:NYMU-Taipei/Project/Time Regulation/Results of Cyanoxilator

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Contents

Extraction of the genomic DNA of S. elongatus PCC 7942

NYMU 20080826 PCC7942 genomic DNA with 1kb marker.JPG

  • tube 1: 101.3 ng/uL (260/280=1.85)
  • tube 2: 72.9 ng/uL (260/280 = 1.90)

PCR of RpaA and SasA

Gel check Comments & Actions
NYMU PCR rpaa sasa.jpg
  1. front part of RpaA
  2. back part of RpaA
  3. front part of SasA
  4. back part ofRpaA
  5. positive control

Expected length(bp)

  1. 389
  2. 420
  3. 750
  4. 472
  5. 1600
NYMU 0910cyanopfu47M.JPG successfully pcr back-SasA
NYMU FRpfu47.JPG successfully pcr front-RpaA
NYMU 0914BR,FS49.jpg
  • comment:

successfully pcr back-RpaA and front-SasA; however they are too weak and we encountered a little bit of trouble doing overlap extension PCR

  • action:
    • gel extraction
    • re-design the primer of FP-in RpaA and RP-in SasA


Reporting assay

Date O.D.600 GFP expression Protocol & Comments
9/29 NYMU 20080929 od m9.jpg NYMU 0929GFPm.jpg
  • The M9 does not seems to have enough nutrition for E. coli to grow.
10/4 NYMU 1004ODm9m.jpg NYMU 1004GFPm9m.jpg
9/30 NYMU 20080930 od lb.jpg NYMU 0930GFPm.jpg * For the 3 hr measure, the plate was not washed carefully, so there is a large background.
10/4 NYMU 1004odlbm.jpg NYMU 1004gfplbm.jpg

Discussion of the reporting

  • Is the freshness of the bacteria important to GFP expression? If so, we should use freshly streaked plates.
  • The white and black plates must be cleaned carefully, otherwise there would be too much background.
  • pBF seemes to grow much faster than the others.

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