General (Stock) Solutions

DNA Gel Sample Loading Buffer—“Blue Juice”
Whiskers
                40 mg Bromophenol Blue                 40 mg Xylene Cyanol
                5 mL Glycerol
                5 mL H­2O
                Vortex, divide into 1 mL aliquots and freeze.

Tris, 1 M pH 7.5
                60.55 g. Trizma Base in 400 mL MQ (Mark’s Quality) H2O                Adjust pH to 7.5 w/ concentrated HCl                q.s. to 500 mL (q.s. = quantum satis/sufficit; as much as suffices)

Tris, 1 M pH 8.0
                60.55 g. Trizma Base in 400 mL MQ (Mark’s Quality) H2O                 Adjust pH to 8.0 w/ concentrated HCl                 q.s. to 500 mL (q.s. = quantum satis/sufficit; as much as suffices)

EDTA, 0.5 M pH 8.0
                60.55 g. Na2EDTA*2H2O (Be sure to check FW before mixing!! Desiccate if necessary.)
                400 mL ddH2O                 Adjust pH to 8.0 w/ solid NaOH                 q.s. to 500 mL

EDTA, 0.5 M pH 8.0
                95.05 g. Na EDTA (FW: 380.2 g.)                 400 mL ddH2O                 Adjust pH to 8.0 w/ solid NaOH                 q.s. to 500 mL

NaCl, 5M
                292.2 g. NaCl in 800 ml MQ H2O                 Stir; adjust to 1 liter total volume.                 pH not important
MgCl2, 1M
                101.65 g. MgCl2*6H2O in 800 mL ddH2O                 Stir; adjust to 1 liter total volume.                 pH not important
TE Buffer, pH 7.4
                10 mL 1M Tris                 2 mL 0.5 M EDTA                 q.s. to 1 L                 check pH and adjust if necessary.
PBS, 10x (Phosphate Buffered Saline)
                40 g. NaCl                 1 g. KCl                 7.2 g. Na2HPO4                 1.2 g. KH2PO4                 450 mL ddH2O                Adjust pH to 7.4 w/ NaOH                 q.s. to 500 mL                 *must be autoclaved or filter-sterilized prior to use.
PBS, 1x (Phosphate Buffered Saline)
                8 g NaCl                 0.2 g KCl                 1.44 g Na2HPO4                 0.24 g KH2PO4                600 mL ddH2O                 Adjust pH to 7.4 w/ HCl.                q.s. to 1 L                 *must be autoclaved or filter-sterilized prior to use.
NaOAc, 3 M (Acetate Solution)

Option 1:
                102.03 g. NaOAc*3H2O                 200 mL ddH2O                 Adjust pH to 5.2 w/ glacial acetic acid                 q.s. to 250 mL
Option 2:
                61.5 g. NaOAc, anhydrous                 200 mL ddH2O                 Adjust pH to 5.2 w/ glacial acetic acid                 q.s. to 250 mL
Ampicillin
                25 mg/mL stock                 Use sterile H2O and filter (0.22 micron)                 Divide into 1 mL aliquots, store at -20oC                 *Becomes yellow when bad
Ethidium Bromide, 10 mg/mL
                0.1 g. Ethidium Bromide                 10 mL ddH2O                 Dissolve by stirring several hours. Cover with foil and store at RT or 4oC
TAE, 50x (Tris-Acetate Buffer with EDTA)
                242 g. Trizma base                 57.1 mL glacial acetic acid                 100 mL 0.5 M EDTA, pH 8                 q.s. to 1 Liter
TBS, 20x (Tris-Buffered Saline)
                48.44 g. Trizma base                 151.9 g. NaCl                 700 mL ddH2O                 Adjust to pH 7.4 with Conc. HCl                 q.s. to 1 Liter
Chloroform:Isoamyl Alcohol (24:1)
                96 mL CHCl3                 4 mL Isoamyl Alcohol                 *mix in hood                 *do not autoclave
Tris (10 mM) EDTA (5 mM) SDS (0.1%)
                2.5 mL 1 M Tris, pH 7.4                 2.5 mL 0.5 M EDTA                 0.25 g SDS                 q.s. to 250 mL with ddH2O
HEPES Buffer, 1M
                238.3 g HEPES (free acid)                 800 mL ddH2O                 Adjust pH w/ NaOH                 q.s. to 1 L with ddH2O
HBSS, 1x  (g/L)

0.185g CaCl2
0.09767g MgSO(anhydrous)
0.4g KCl
0.06g KH2PO4 (anhydrous)
0.35g NaHCO3
8.0g NaCl
0.04788g Na2HPO4 (anhydrous)
1.0g D-Glucose
q.s. to 1L with ddH2O
*sterile filter before use

INS -1  Supplement
100mL  Glutamine solution (200mM)
100mL  Na-pyruvate solution (100mM)
35.2uL  2-mercaptoethanol

RIPPA Buffer
10 mM Tris Cl (1 mL of a 1M solution)
1mM EDTA (200 uL of a 0.5M solution)
0.5mM EGTA (100ul of a 0.5M solution)
1% Triton x 100 (10 mL of a 10% solution)
0.1% Sodium Deoxycholate (1mL of a 10% solution)
0.1% SDS (1mL of a 10% solution)
140mM NaCl (2.8mL of a 5M solution)
- combine all above liquids then q.s. to 100 mL