Team:NTU-Singapore/Notebook/30 May 2008

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Contents

Friday 30 May 2008

  • Hung (Greenbear) and Choon Kit: Analysis of Fe promoter plasmid by running Gel Electrophoresis (the remaining steps of phase III).

Experiment No 3

Date 27-28 May 2008 
Start Time 1030						
End Time 1130

Personnel:

Choon Kit, Hung, Dr Tan TL

Title of Experiment:

Analysis of Fe promoter plasmid by running Gel Electrophoresis

Materials:

Pipette with pipette tips
DNA marker (10kb)
DNA marker (1000pb)
Blue loading dye
Gel Electrophoresis setup
PCR tube with ligated samples of Fe promoter Biobricks

Protocols/Procedures:

  1. Prepare the agarose gel setup. Add 2ul of blue loading dye to 10ul of ligated sample in the PCR tube. Load 10ul of 10kb DNA marker, 12ul of ligated plasmid sample 1 and 2, and 10ul of 1000bp DNA marker into each electrophoresis well.
  2. Cover the gel electrophoresis, ensuring the wires are plugged in place. Turn on and run the electrophoresis for 15min at 120V, ensuring the loading dye does not exceed the whole span of agarose gel.
  3. Scan the agarose gel under UV.

Observations:

Gel electrophoresis indicated bands between 300 – 350bp and 2000bp. 350bp corresponded to the expected length of Fe promoter. 2000bp corresponded to the length of the ligated plasmid.


Conclusion:

Preliminary test indicated that the Miniprep was successful.

Notes:

Refer to video clip 30 May 08 for details.