Merlin Preps Reagents (1/3)

Chia Jin Ngee mcblab47 at leonis.nus.sg
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)

PS1
50 mM Tris.Cl, pH7.6
10 mM EDTA
100 mg/ml RNAse A

PS2 
0.2 M NaOH
1% SDS

PS3
61.35 g potassium acetate
35.7 ml glacial acetic acid
MilliQ water to 500 ml

PS4
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

PS5
200 mM NaCl
20 mM Tris.Cl, pH 7.6
5 mM EDTA
50% ethanol (Aristar grade)

TE buffer
10 mM Tris.Cl, pH 8.0
1 mM EDTA

MPS4
Same as PS4 but with 15 g of Celite 545

MPS5
Same as PS4 but without Celite 545

--
Jin Ngee, Chia			Chemical Carcinogenesis Laboratory
(Genie, the OligoMan)		Institute of Molecular and Cell Biology
mcblab47 at leonis.nus.sg		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



More information about the Methods mailing list