Maheshri:Buffers
50X TAE
For 500mL:
1. Dissolve 121g of Tris in 350mL of water.
2. Add 28.6mL of Glacial acetic acid
3. Add 50mL of 0.5M EDTA
20X SSC
For 1L:
1. Dissolve 175.3g of NaCl, 88.2g of sodium citrate in 900mL of water
2. pH to 7.0 with HCl
3. Mix
10% SDS
For 1L:
1. In a chemical fume hood, dissolve 100of SDS in 1L of water
2. Filter sterilize if needed
0.5M EDTA
For 1L:
1. Dissolve 186.10g of EDTA Disodium salt in 700mL of water
2. Adjust pH to 8.0 with NaOH. EDTA won't dissolve unless the pH is right.
3. QS to 1L
1M Tris pH 7.9
For 1L:
1. Dissolve 121.1g Tris in 700mL water
2. Adjust to pH 7.9 with HCl (approx 45mL)
3. QS to 1L
5M NaOH
For 1L:
In a chemical fume hood dissolve 400g of NaOH in a total of 1L
5X SDS-PAGE Buffer
For 1L:
1. Dissolve 15.1g of Tris base, 94g of Glycine, and 5g of SDS in 900mL of water
2. QS to 1L
Transfer Buffer for Western Blots
5X stock:
Tris base 15.15g
Glycine 72g
Water to 1L, store at room temperature
1X working solution (mix on the day of use):
5X stock 200mL
Water 500mL
Methanol 300mL
Chill at 4°C before use
Stripper for Western Blots
Acetic Acid 15mL
NaCl 14.6g
Water to 500mL, store at room temperature