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Lab July 2 2009
From 2009.igem.org
Thursday, July 2, 2009
got the pH meter and drychem for remaining stocks
today intending to make 1L TE, Tris and EDTA stock solutions, and 1L of P1
1L of 1M Tris-HCl at pH 8.0
121.14 g Tris base in 500 mL water
pH adjust with HCl to pH 8.0, top to 1L
500 mL of 1M Tris-HCl at pH 7.5
60.57 g Tris base in 250 mL water
pH to 7.5 and top up to 500mL
1L of PB buffer @ 5M Gu-HCl, 30% ethanol
477.65 g of Gu-HCl
315.79 ml ethanol from 95% ethanol stock
topped up to 1L with dH2O
1L of N3 buffer @ 4.2M Gu-HCl and 0.9M potassium acetate, pH 4.8
401.23 g Gu-HCl
88.33 g KCH3COO
add HCl to adjust pH
top up to 1L
Rather than talking about the buffers as P1, P2, N3, PB, PE, and EB, Dr. Nano would prefer we call them by functional names. (the following is just my cut at the names - please replace with better ones if you have more consise names for these! --Derek)
P1 = alkaline prep buffer
P2 = lysis buffer
N3 = neutralizing buffer
PB = endonuclease wash
PE = ethanol wash
EB = elution buffer (.1xTE)
Also, we are to limit questions to Dr. Nano to between the top of the hour and quarter after the hour. He needs uninterrupted time to get some writing done.
Final production for the day:
1L Tris-HCl @ pH 8.0
500 mL Tris-HCl @ pH 7.5
1L PB // endonuclease wash
1L N3 // neutralizing buffer
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