C.B.S. Scientific TTGEK-2401-220 - Manual (Page 51)

TTGE Instruction Manual, version 8/30/2011
51
www.cbsscientific.com
SECTION 4
Running Conditions
4.1 Recommended Power
1. 150V (40mA/one dual cassette) CV for 5-7 hours
4.2 Recommended Buffers
Warning: Do not use deionized water with a specific resistance > 1 meg ohm.
STOCK SOLUTIONS
Acrylamide Stock Solution 40% Acrylamide/Bis (37.5:1)
For 100ml use the following:
38.93 g of acrylamide
1.07 g of Bis-acrylamide
Add dH
2
O to 100ml. Do not autoclave
50x TAE Gel Running Buffer
For 1 liter use the following:
242g Tris Base
57.1ml Glacial acetic acid
100ml .5 M EDTA, pH 8.0
Add dH
2
O to 1 liter
Denaturant Stock Solution (7.5% gel)
For 100ml use:
10% 20% 30% 40% 50% 60% 70% 80% 90% 100%
40% Acrylamide/Bis (ml) 18.8 18.8 18.8 18.8 18.8 18.8 18.8 18.8 18.8 18.8
50x TAE Buffer (ml) 2 2 2 2 2 2 2 2 2 2
Formamide (ml) 4 8 12 16 20 24 28 32 36 40
Urea (g) 4.2 8.4 12.6 16.8 21 25.2 29.4 33.6 37.8 42
Add dH
2
0 to 100ml
Store denaturant stock solutions at 4º C in amber bottles
Polymerization Catalysts:
1. TEMED
2. APS Stock Solution
For 50 ml of APS stock (10%) use 5 g of ammonium persulphate and water to 50ml. Store at 4ºC.
Neutral Loading Solution
For preparation of 10ml:
20% Ficoll or sucrose, 2 g of Ficoll 400 or sucrose
10mM Tris-HCl, pH 7.8, 100ul of 1.0 M Tris-HCl, pH 7.8
1mM EDTA, 20 ul of 0.5 M EDTA, pH 8.0
0.1% dye (BPB), 10 mg of dye
Water to 10ml