Team:Illinois/Bimolecular Fluorescence Biosensor Notebook

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1st July

TE Buffer Recipe
- Uses
  - TE used to bring up oligos into solution

- People who know how to do this
  - Joleen, Luke

- Concentration
  - 10mM Tris
  - 1mM EDTA
  - pH 8.0

- Method
  - For 500mL volume
    - Add: 0.1861g EDTA; 0.6057g Tris to 1 liter flask
    - Bring it up to 500mL with Deionized water (filtered in ---? container)
    - Put on "corning" mixer; get a larger stir bar from drawer and put on slow ~20 RPM rotation; no heat for 2-3 minutes until fully dissolved.
    - Standardize the pH meter (instructions on sign at prep bench)
    - Put gloves on; put pH electrode in flask but first add stir bar and put on low speed mixing
    - Bring pH to 8.0 (i.e. 8.00 +/- 0.05) by adding base (NaOH) or acid (HCl) in very small drops using a plastic pipette. (Wait for pH meter to eqiullibrate after each drop)

IMP: Put cap back on bottom of pH probe

- - - Put coloured tape along side of flask and label with pH, name and iGEM
    - Cover it with heavy duty aluminium foil (just like a square inch to cover the top) and put a strip of autoclave tape along top of foil
    - Optional: out in big plastic autoclave bin
    - Bring to autoclave room; do not use the big "Beta Star"

    Set on: 15 minutes; ~250 degrees Farenheit = ~121 degree Celcius; "liquid" run; for "operator #" just press enter; then "Run"; tubes = ~ 45 minutes; total to run since it must cool down and decompress

- - - (Some extra side notes still to add)

TAE Buffer Recipe

TBE Buffer Recipe
- - 1 liter of 5x TBE Running Buffer, pH 8.13-8.23

- Materials
  - Tris-base - 54.0g
  - Boric Acid - 27.5g
  - EDTA - 2.92g
  - DI Water - 1.0L

- Method
  - Stirred with stirring rod for 3-5 minutes