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wiki:documentation:wiki:documentation:jun:03-12-2014 [2019/12/04 08:57] (current)
jliu created
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 +====== 03-12-2014 ======
 +
 +I: worm freezing \\ 
 +
 +The following worms are frozen. 2X10cm plates of worms are washed off with ~6ml of freezing solution. Then 0.5ml of suspended worms are added into 10X cryotubes. 1 of them are kept as thawing control. \\ 
 +
 +1: BC11358 \\ 
 +2: ZW284 [note: this strain carries an extrachromosomal array. There were enough GFP worms on the plates before freezing] \\ 
 +3: HW2678 \\ 
 +4: HW2679 \\ 
 +5: AML328 (7 tubes frozen as this is just a supplement for previous freezing). \\ 
 +
 +If the freezing is successful, then all the strains from Monika and me are frozen (except N2, underway). The ones recently received from CGC are also to be frozen once ready. \\ 
 +
 +II: make solutions \\ 
 +
 +Since the worms seem to grow slower. We want to rule out whether this is due to the solutions and NGM made by Azubis. We decide to make all solutions by ourselves. Nicoline made the potassium phosphate solution and I made the rest. \\ 
 +
 +1) Cholesterol 5 mg ml-1 in 95% EtOH (100ml) [1.1g Cholesterol in 200ml of EtOH] \\ 
 +2) 1M CaCl2 (200ml) [22.2g of CaCl2] \\ 
 +3) 1M MgSO4 (200ml) [49.3g of MgSO4.7H2O]. Note: the MgSO4 in our lab contains 7H2O, pay attention to the MW. \\ 
 +
 +4) NGM media \\ 
 +3 g NaCl \\ 
 +17 g agar \\ 
 +2.5 g peptone \\ 
 +1 ml cholesterol (5 mg ml-1 in 95% EtOH) \\ 
 +975 ml H2O \\ 
 +
 +Autoclave, and then add the following sterile solution (autoclaved) \\ 
 +1 ml 1 M CaCl2 \\ 
 +1 ml 1 M MgSO4 \\ 
 +25 ml 1 M potassium phosphate (pH 6) (to avoid precipitation,​ mix between addition of MgSO4 and potassium phosphate. \\ 
 +
 +5) Freezing solutions (I know this one made by Julius worked as the freezing worked well. Since it is getting low, I made 500ml) \\ 
 +Recipe: ​ \\ 
 +NaCl 2.9g \\ 
 +1M KH2PO4 (pH6.0) 25ml \\ 
 +Glycerol 120ml \\ 
 +H2O 355ml \\ 
 +
 +Will add 150ul of 1M MgSO4 to it after autoclaving. \\ 
 +
 +
  
wiki/documentation/wiki/documentation/jun/03-12-2014.txt ยท Last modified: 2019/12/04 08:57 by jliu

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