TE (100:10) buffer: 100 mM Tris-HCl, pH 7.6, 10 mM EDTA

                       100 ml                       1 M Tris-HCl, pH 7.6

                       20 ml              0.5 M EDTA

                       ddH2O to 1 L

TE (50:1) buffer: 50 mM Tris-HCl, pH 7.6, 1 mM EDTA

                       0.5 ml             1 M Tris-HCl, pH 7.6

                       0.1 ml             100 mM EDTA

                       9.4 ml             ddH2O

                       10 ml

TE-RNase solution: 50:10 TE buffer containing 40 ug/ml RNase A and 40 U/ml RNase T1

                       1.2 ml             1 M Tris-HCl, pH 7.6

                       480 ul             0.5 M EDTA

                       50 ul               20 mg/ml RNase A

              10 ul           100U/ul RNase T1

                       22.3 ml           ddH2O

                       24.0 ml

Tetracycline stock (Tet): Stock of 10 mg/ml in 50% ethanol + sddwater.

                       1 g                   Tetracycline (Sigma T-3383)

                       50 ml              100% ethanol

                       sddH2O to 100 ml (store at 4deg.C in the absence of light)

Add to media for final conc. 20 ug/ml.

1% thiamine:

                       100 mg           thiamine (Sigma T-4625)

                       sddH2O to 

                       10 ml (filter sterilized)

1M Tris-HCl, pH 7.6, 8.0, 8.5, 9.0, 9.5:

                       121.1 g           Tris base

                       ddH2O to 800 ml

Adjust pH with concentrated HCl and then add ddH2O to 1 L.

10X TM buffer: 500 mM Tris-HCl, pH 8.0, 150 mM MgCl2 in sterile double distilled water.

                       5 ml                 1 M Tris-HCl, pH 8.0

                       1.5 ml             1 M MgCl2

                       sddH2O to 10 ml

50:2:10 TTE: 50 mM Tris-HCl, pH 8.0, 2% Triton X-100, and 10 mM EDTA in double distilled water.

                       5 ml                 1 M Tris-HCl, pH 8.0

                       2 ml                 0.5 M EDTA

                       2 ml                 Triton X-100 (Sigma X-100)

                       ddH2O to 100 ml

TTE: 10 mM Tris-HCl, pH 8.0, 0.5% Triton X-100, and 0.1 mM EDTA in double distilled water.

                       500 ul             1 M Tris-HCl, pH 8.0

                       250 ul             Triton X-100 (Sigma X-100)

                       10 ul               0.5 M EDTA

                       ddH2O to 50 ml

X-gal (5-bromo-4-chloro-3-indolyl b-D-galactopyranoside):

                       200 mg           x-gal (Sigma B-4252)

                       dimethylformamide 

                       (DMF) to 10 ml

                       Aliquot and store protected from 

                       light at -20degC)

2xTY medium:

                       16 g                Bacto-tryptone (Difco 0123-01-1)

                       10 g                Bacto-yeast extract (Difco 0127-05-3)

                       5 g                   NaCl

                       ddH2O to 1 L (autoclave)