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Bio Ecology Essay

284 words - 2 pages

Gel Electrophoresis Protocol

Materials:

40 ml of 1% agarose gel
40 ml of TBE 1x buffer
4 microL of Ethidium Bromide
50 ml graduated cylinder
250 ml Erlenmeyer flask
Ladder
*necessary aliquots
Gel Cast and comb
Mini sub cell chamber
Goggles
Heat protecting gloves

Procedure:
1. Weigh 0.40 g agarose gel
2. Obtain 40 ml of TBE 1x buffer into a 50 ml graduated ...view middle of the document...

Cover Erlenmeyer flask with kin wipe
5. Microwave until the mix bubbles vigorously at about 30-40 seconds
6. Remove mix from microwave using heat protecting gloves and allow to cool down for about 5 minutes
7. Sterilize the gel cast with 70% ethanol and balance the cast accordingly
8. Add 4 microL of ethidium bromide into the mix then swirl
9. Pour the mix into the gel cast (pop air bubbles with a pipettor tip)
10. Sterilize comb with ethanol and put it in the gel cast
11. Cover the gel with kin wipe and wait for 30 minutes (if in a hurry put the apparatus in the fridge for 15 mins)
12. Remove the comb slowly and gently
13. Remove the gel and transfer it into a mini sub cell chamber with the wells on the black side
14. Pipet the ladder into the farthest well then the rest of the aliquots accordingly
15. Cover the mini sub cell, attach the wires and change the machine setting to 100volts
16. Run machine for 1 hour
17. Remove gel from the apparatus, bring to the UV light room, take pictures of the gel and save them

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