Merlin Preps Reagents (1/3)

Chia Jin Ngee mcblab47 at
Fri Feb 18 03:03:30 EST 1994

Merlin Preps Reagents
(see: Merlin Minipreps, Merlin Maxipreps)

Reagents needed:

1 M Tris.Cl, pH 7.6
0.5 M EDTA, pH 8.0
2 M NaOH (stored in plastic bottles, silicates from glass bottles interfere with yield)
10% SDS
potassium Acetate
glacial Acetic Acid
guanidine hydrochloride (also known as guanidinium/guadinium chloride/hydrochloride)
diatomaceous earth-flux calcined (also known as Celite 545, Fluka cat no 22140)
RNAse A (10mg/ml DNAse free)

50 mM Tris.Cl, pH7.6
10 mM EDTA
100 mg/ml RNAse A

0.2 M NaOH
1% SDS

61.35 g potassium acetate
35.7 ml glacial acetic acid
MilliQ water to 500 ml

66.84 g guanidine hydrochloride
33.333 PS3
Use mild heat to dissolve, will not dissolve completely but some fine precipitate will remain
Adjust to pH 5.5 using 10 M NaOH (usually 1.5 - 2.0 ml)
MilliQ water to 100 ml (precipitate will dissolve now)
Filter the solution with a 0.45 mm filter
Add 1.5 g of Celite 545 to the solution
Shake well and aliquot to two 50 ml conical centrifuge tubes (Falcon)
Wrap with aluminium foil and store at room temperature

200 mM NaCl
20 mM Tris.Cl, pH 7.6
50% ethanol (Aristar grade)

TE buffer
10 mM Tris.Cl, pH 8.0

Same as PS4 but with 15 g of Celite 545

Same as PS4 but without Celite 545

Jin Ngee, Chia			Chemical Carcinogenesis Laboratory
(Genie, the OligoMan)		Institute of Molecular and Cell Biology
mcblab47 at		National University of Singapore
Tel: (065)772-3797		Kent Ridge Crescent
Fax: (065)779-1117		Singapore 0511
"I'm Mr Chia not Ms Chia and I know Genie is a girl's name"-Jin Ngee, Chia

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