Department of Physics, Chemistry and Biology (IFM)

Shorty Buffer

0.2 M Tris-HCL, pH 9.0

0.4 M LiCl

25mM EDTA, pH 8.0

1% SDS

H₂O

 

TE Buffer

10 mM Tris, pH 8.0

1 mM EDTA, pH 8.0

H₂O

PCR mix:

5 µl 10×Paq5000 reaction buffer

0.4µl 100mM dNTP

1µl 10µM forward primer

1µl 10µM reverse primer

100ng genomic DNA

0.5µl of Paq5000 DNA Polymerase

Gel preperation (0.8%)

200 ml TBE

1.6 mg agarose

2 µl Ethidium bromide

Sample preparation

8µl sample

2µl loading dye

 

Isolation buffer

50 mM Tricine (pH 8)

0.3 M Sorbitol

1 mM EDTA

2 mM MgCl2

Add BSA freshly when needed

Seperation buffer (pH 8.83)

1.5 M Tris-HCL

0.4 % SDS

dd H₂O

Stacking buffer (pH 6.8)

0.5 M Tris HCL

0.4 % SDS

dd H₂O

Running buffer (×10) (pH 8.3)

0.25 M Tris

1.9 m glycin

1 % SDS

dd H₂O

Before use, dilute to 10 times.

Solubilization buffer, S ×3 (pH 6.8)

0.5 M Tris-HCL (pH 6.8)

0.4 % SDS

87 % glycerol

dd H₂O

Before use, mix 417 µl S×3, 83 µl 600 mM DTT (Dithiotreitol) and BPB.

TEMED (N’,N’,N’,N’- tetramethyl-ethylenediamine)

10 % APS (Ammonium persulphate)

APS

dd H₂O

Blotting buffer

48 mM Tris

39 mM glycin

0.0375 % SDS

20 % methanol

dd H₂O

TBS×5 (Tris buffered saline) (pH 7.5)

100 mM Tris-HCL

0.5 M NaCl

dd H₂O

Prepare TBS×1 by diluting it 5 times.

Washing/Antibody solution (TTBS×1) (pH 7.5)

TBS×1

0.1   % Tween-20

Blocking solution (pH 7.5)

TTBS×1

0.5 % non-fat dry milk

Antibody 1 (ab 1)

Rabbit IgG raised from the desired protein.

Antibody 2 (ab 2)

Donkey anti-rabbit IgG horseradish peroxidase conjugate (HRP)

ECL-Plus Chemiluminescent substrate

1 ml of solution A

25 µl of solution B

1 ml dd water.