Team:MIT/Protocols

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Protocols

Cloning	Tissue Culture	Miscellaneous Lab	Non-Lab
Gels			MIT iGEM cookie preparation

Gels

Preparing the Gel Dissolve UltraPure agarose to a final concentration of 1%(by mass) in TAE buffer in a glass bottle. Heat the solution in the microwave with frequent stirring to dissolve the agarose homogenously. ~1 minute/200ml solution Place the solution in a warm water bath for 5 mins. Add 10 µl SYBRSafe (1:10000) per 100 ml of the solution and mix well. Pour 50ml of solution per small gel tray. (the gel trays and combs should be pre-cleaned with water and wiped dry). -Note for combs: 15-well combs hold about 6 ul liquid, 12-well combs hold about 15 ul, 8-well combs hold about 20 ul -Taping two 8-well comb wells together results in a well that holds up to 100 ul -Taping three 8-well comb wells together result in a well that holds up to 200 ul Use 120ml per large gel tray. [need to update amounts] For the small set: small trays hold 20ml, large trays hold 50ml Wait for the gels to solidify. ~15 mins Label and store at 4C.

MIT iGEM cookie preparation

Reagents required:
1 1/2 cups (3 sticks) unsalted butter
1 cup sugar
2 large egg yolks
3 3/4 cups sifted all-purpose flour
1/4 teaspoon salt
1 tablespoon pure vanilla extract

Safety note: wear gloves, this procedure involves touching the dough with your hands, a lot. Human tissue is minimum BSL 2 because it can contain contagious pathogens.
Make sure oven is set to 350°F
Melt 3 sticks butter
Add butter and 1C sugar to mixing bowl
mix sugar and butter thoroughly
Add 3.75c all purpose flour
Add .25 tsp salt
Add 1.33Csugar
Add 0.75c cocoa powder
Add 1Tbsp vanilla extract
Add 2 egg yolks
mix until consistent and cohesive, this will likely require kneading.
for all dough
place some amount of dough onto cookie sheet and flatten with palm of hand, should be >3" in diameter and ~.25" thick and flat (no texture/bumps)
press outline cutter into dough and apply pressure around perimeter to ensure a good cut
--The dough should just make it to the angled part of the cookie cutter, if not, adjust height on next cookie
remove cookie cutter
removed excess dough
get more dough and repeat cookie blanking until cookie sheet is full
press stamp into each cookie by pushing on two opposite edges, examine result to see if enough pressure was applied. A cookie can be stamped twice if you're careful.
bake cookies on sheet for 9 minutes
(at this point, running multiple parallel sheets is suggested)
after baking, use a spatula to move cookies onto drying rack
--This is the easiest step to break cookies at, they almost always break teeth off the gears. Make sure that cookies are rotated and aligned such that all gear teeth are supported and they do not fall in between wires on rack.
Wait as long as possible for cookies to cool completely before transferring to a storage container as cookie structural integrity is inversely proportional to temperature. Having a fan blow on cookies to help cooling is suggested.