Difference between revisions of "20.109(F16):Prepare comet chips (Day1)"
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#Use the ruler in your team drawer to draw a X x X rectangle on the gelbond film. | #Use the ruler in your team drawer to draw a X x X rectangle on the gelbond film. | ||
#*Divide the rectangle into four quadrants and label as depicted in the image on the right. | #*Divide the rectangle into four quadrants and label as depicted in the image on the right. | ||
| − | #Prepare 20 mL of 1% normal melting point (NMP) agarose. | + | #Prepare 20 mL of 1% normal melting point (NMP) agarose. <br>Be careful as the agarose solution will be very hot!<br> |
#*Calculate the amount of NMP agarose powder needed for a 1% w/v solution. Check your math with an instructor before you continue. | #*Calculate the amount of NMP agarose powder needed for a 1% w/v solution. Check your math with an instructor before you continue. | ||
#*Obtain a small milk bottle from the front bench. | #*Obtain a small milk bottle from the front bench. | ||
Revision as of 15:32, 29 June 2016
Contents
Introduction
comet chip assay...
Protocols
Part 1: Laboratory orientation quiz
Complete the orientation quiz with your partner. Though you are working with your partner, each student should record all answers on the provided quiz. If you disagree with your partner on an answer, you should write what you think is the correct answer on your quiz.
Good luck!
Part 2: Prepare comet chips
blah...
- Obtain a sheet of gelbond film from the laboratory bench at the front of the room. The paper is on the hydrophilic side of the gelbond film.
- Be sure to keep the paper associated with the gelbond film so you know which side is which.
- Use the ruler in your team drawer to draw a X x X rectangle on the gelbond film.
- Divide the rectangle into four quadrants and label as depicted in the image on the right.
- Prepare 20 mL of 1% normal melting point (NMP) agarose.
Be careful as the agarose solution will be very hot!
- Calculate the amount of NMP agarose powder needed for a 1% w/v solution. Check your math with an instructor before you continue.
- Obtain a small milk bottle from the front bench.
- Weigh out the appropriate amount of NMP and add it to the milk bottle.
- Use a cylinder to measure 20 mL of 1x PBS and add it to the milk bottle with the NMP agarose powder.
- Swirl to mix.
- To melt the NMP agarose, microwave the solution for 20 sec intervals. After each interval, remove the milk bottle and gently swirl while checking for unmelted agarose crystals.
- When no crystals are visible in the solution take the milk bottle to your bench.
Part 3: Split mammalian cells
Reagents
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