Team:Michigan/Media Recipes
From 2010.igem.org
(Difference between revisions)
(17 intermediate revisions not shown) | |||
Line 7: | Line 7: | ||
</style></html> | </style></html> | ||
+ | |||
+ | {|cellspacing=0 | ||
+ | |-valign="top" | ||
+ | |width="700px" style="background-color:rgb(190,210,250);"| | ||
+ | |||
==Media Recipes== | ==Media Recipes== | ||
+ | |||
+ | [[Team:Michigan/Media Recipes#General Recipes|General Recipes]] | ||
+ | |||
+ | [[Team:Michigan/Media Recipes#Oil Sands Recipes|Oil Sands Recipes]] | ||
+ | |||
+ | ==General Recipes== | ||
'''LB media''' | '''LB media''' | ||
- | *10 g LB | + | *10 g LB medium |
*500 mL DI water | *500 mL DI water | ||
''autoclave to sterilize'' | ''autoclave to sterilize'' | ||
'''LB media with agar''' | '''LB media with agar''' | ||
- | *10 g LB | + | *10 g LB medium |
*7.5 g agar | *7.5 g agar | ||
*500 mL DI water | *500 mL DI water | ||
Line 61: | Line 72: | ||
*1049 uL of DI water | *1049 uL of DI water | ||
''filter sterilize with .22 um filter'' | ''filter sterilize with .22 um filter'' | ||
+ | |||
+ | '''40% glucose solution''' | ||
+ | *20 g of glucose | ||
+ | *50 mL of DI water | ||
+ | ''filter sterilize'' | ||
+ | |||
+ | '''0.1 M NaOH solution ''' | ||
+ | |||
+ | BE VERY CAREFUL WHEN MIXING! SOLID NaOH BURNS YOUR SKIN BUT YOU MAY NOT FEEL IT AT FIRST! MAKE SURE ANY SPILLS ARE CLEANED UP VERY WELL! | ||
+ | *0.2 g of NaOH | ||
+ | *50 mL of sterile water | ||
+ | ''Filter Sterilize'' | ||
+ | |||
+ | '''10mM dNTP mix for PCR''' | ||
+ | |||
+ | ''MAKE SURE ALL REAGENTS ARE WELL MIXED BEFORE STARTING BY PULSE CENTRIFUGING OR FLICKING'' | ||
+ | *100 uL of 100mM dATP | ||
+ | *100 uL of 100mM dTTP | ||
+ | *100 uL of 100mM dGTP | ||
+ | *100 uL of 100mM dCTP | ||
+ | *600 uL of ultra pure water | ||
+ | ''aliquot into 50 uL volumes (this will take 20 1.5mL eppendorf tubes)'' | ||
+ | |||
+ | ==Oil Sands Recipes== | ||
'''1.5 mg/mL Cyclohexanecarboxylic acid solution''' | '''1.5 mg/mL Cyclohexanecarboxylic acid solution''' | ||
- | Cyclohexanecarboxylic acid is volatile should only be | + | Cyclohexanecarboxylic acid is volatile and should only be used under a hood. For accurate measurement, use a covered scale with sliding doors (in the hood). Store in the corrosive cabinet. |
*0.075 g cyclohexanecarboxylic acid | *0.075 g cyclohexanecarboxylic acid | ||
Line 76: | Line 111: | ||
*0.002 g of FeCl3 | *0.002 g of FeCl3 | ||
*318 mL of DI H2O | *318 mL of DI H2O | ||
- | Autoclave at 15 psi and | + | ''Autoclave at 15 psi and 121°C for 30 minutes'' |
After the media has cooled completely add: | After the media has cooled completely add: | ||
*665 uL of 1 M MgSO4 solution | *665 uL of 1 M MgSO4 solution | ||
*72 uL of 1 M CaCl2 solution | *72 uL of 1 M CaCl2 solution | ||
- | |||
- | |||
- | |||
- | |||
- | |||
'''Bushnell-Haas Media with glucose''' | '''Bushnell-Haas Media with glucose''' | ||
Line 107: | Line 137: | ||
*2 mL of sterile DI water | *2 mL of sterile DI water | ||
*6.25 mL of casamino acid solution | *6.25 mL of casamino acid solution | ||
+ | |||
+ | '''Bushnell-Haas Media with cyclohexanecarboxylic acid adjusted to pH of 9''' | ||
+ | *39.75 mL of Bushnell-Haas Salts | ||
+ | *4 mL of cyclohexanecarboxylic acid solution | ||
+ | *6.25 mL of sterile DI water | ||
+ | Test how much 0.1 M NaOH must be added to 1 mL of media to raise it to a pH of 9 (add 1 uL of base at a time) | ||
+ | |||
+ | Scale up the amount of 0.1 M NaOH solution to add to the remaining volume of media (approaching final amount slowly) | ||
+ | |||
+ | On 8/3/2010, approximately 20 uL of 0.1 M NaOH had to be added to 1 mL to get to a pH between 8 and 9 (tested with pH paper) and 675 uL of 0.1 M NaOH was added into the 45 mL of remaining media. | ||
+ | |||
+ | |width="250px" style="background-color:rgb(190,210,250)"| | ||
+ | |||
+ | [[Image:Lab01.png|middle|250px]] | ||
+ | |||
+ | [[Image:Lab02.png|middle|250px]] | ||
+ | |||
+ | [[Image:Lab04.png|middle|250px]] | ||
+ | |||
+ | [[Image:Lab05.png|middle|250px]] | ||
+ | |||
+ | [[Image:Lab06.png|middle|250px]] | ||
+ | |||
+ | [[Image:Lab09.png|middle|250px]] | ||
+ | |||
+ | [[Image:Lab07.png|middle|250px]] | ||
+ | |||
+ | [[Image:Lab08.png|middle|250px]] | ||
+ | |||
+ | [[Image:Lab10.png|middle|250px]] | ||
+ | |||
+ | [[Image:Lab11.png|middle|250px]] | ||
+ | |||
+ | |} |
Latest revision as of 23:52, 26 October 2010