Lab July 2 2009

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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