**2.7 Testing for protein expression and solubility in** *E. coli*

1.LB.

*Growing and Handling of Bacterial Cultures*

8.LB-agar plates containing the appropriate antibiotic.

1.Transformed colonies on LB-agar plate (see Section 3.3).

**2.3 Calculating efficiency of competent cells**

6.Sterile aluminum foil or culture tube cap.

raacetic acid (EDTA), 20 μg/mL RNase A.

6.TE buffer: 10 mM Tris-HCl, pH 8.0; 0.1 mM EDTA.

a.Make the 50% glycerol solution by diluting 100% glycerol into water.

1.Resuspension buffer: 50 mM Tris-HCl, pH 8.0; 10 mM ethylenediaminetet-

2.Lysis buffer: 200 mM NaOH, 1% w/v sodium dodecyl sulfate (SDS).

3.Precipitation buffer: 3 M potassium acetate, 2 M glacial acetic acid, 4°C.

1. 50% glycerol solution (autoclaved).

2.Screwtop cryogenic vials.

**2.6 DNA plasmid purification**

4.Wash buffer: 70% ethanol.

5.95% (or 100%) ethanol.

3.Liquid nitrogen.

**2.4 Inoculating overnight cultures**

2. 15 mL conical tube.

3.Sterile inoculating loop.

4.Appropriate antibiotics.

5.Shaker/incubator.

**2.5 Glycerol stocks**

7.Competent cells.

9.Plasmid DNA.

10.Heat block set.

1.LB.

**40**

