General (Stock) Solutions
DNA Gel Sample Loading Buffer—“Blue Juice”
- 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.
DPBS, 1x, With Calcium and Magnesium (Based on Biological Industries Fromulation
- 49.95mg CaCl2
- 100.125mg KCl
- 99.96mg KH2PO4 (Monobasic)
- 23.80mg MgCl2
- 4g NaCl
- 575mg Na2HPO4 (Dibasic)
- q.s to 450mL
- pH to 7.4
- q.s to 500mL
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 MgSO4 (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