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Study Notes: SDS-PAGE Handy Hints

Initially the use of the SDS-PAGE is confusing and the assembly and gel preparation steps are complicated and fiddly. There is no substitute for hands-on experience but there are many common sense approaches that will make the use of the SDS-PAGE easier with fewer mistakes. We call these’ Handy Hints’.

Handy Hints for using the virtual SDS-PAGE

  1. Print out a copy of the SOP: SDS-PAGE and have this on-hand.
  2. Read the SOP very carefully before commencing.
  3. When working with the ‘virtual’ SimuLab SDS-PAGE read the screen instructions and other information very carefully and take note of important points.
  4. Follow the on-screen instructions exactly as stated.
  5. Be sure to use the parameters listed in the SOP and check/adjust the SDS-PAGE to reflect these parameters. The parameters that will need to be adjusted are:
    • add 20 µL of sample to well
    • turn on stirrer base
    • turn on powerpack and set to 10 mA
    • turn gel off when dye front is one cm from the bottom
    • fix gel for one hour
    • stain gel for two hours
    • destain gel for two hours
    • drying apparatus - apply vacuum, set temperature to 60°C and dry for two hours.
  6. Print out the images of the gel(s) for future reference and remember to obtain the densitometer results for the concentration of acceptable sample of HGH.
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