DNA Isolation
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
H2OPCR
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
Agarose gel electrophoresis
Gel preperation (0.8%)
200 ml TBE
1.6 mg agarose
2 µl Ethidium bromide
Sample preparation
8µl sample
2µl loading dye
Isolation of root plastids
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.
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Director of undergraduate studies Biology
Last updated:
05/20/09