**2.6 DNA plasmid purification**


**41**

lysozyme.

*Growing and Handling of Bacterial Cultures within a Shared Core Facility for Integrated…*

1.Induced cells suspended in lysis buffer with a protease inhibitor cocktail, 0.1 mg/mL DNase I, 1 mg/mL lysozyme and 0.1 mg/mL 4-(2-Aminoethyl)

3.Sample loading buffer: 10% glycerol, 0.14 M Tris Base, 0.1 M Tris-HCl, 2% lithium dodecyl sulfate (LDS), 0.5 mM EDTA, 0.02% Blue G250; 0.006%

4.Running buffer: 50 mM 2-(N-morpholino)ethanesulfonic acid (MES), 50 mM

1.Buffer A: 50 mM Tris pH 7.5, 100 mM NaCl, 5 mM EDTA, 1 mg/mL lysozyme.

2.Buffer B: 50 mM Tris pH 7.5, 2 M NaCl, 5 mM EDTA, 1 mg/mL lysozyme.

3.Buffer C: 50 mM Tris pH 7.5, 100 mM NaCl, 50% detergent, 1 mg/mL

*DOI: http://dx.doi.org/10.5772/intechopen.81932*

2.Appropriate antibiotics.

4.Microcentrifuge tubes.

3.Incubator/shaker.

2.Sonication buffer.

3.Ice-saltwater bath.

1.Electrophoresis system.

2.4–12% Bis-Tris mini gel.

5.Coomassie Blue stain.

6.Protein molecular weight marker.

**2.10 Testing lysis conditions for solubility**

5.Centrifuge.

**2.8 Lysing cells**

1.LB.

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

6.Isopropyl β-D-1-thiogalactopyranoside (IPTG).

benzenesulfonyl fluoride (AEBSF).

4.Probe sonicator equipped with ½ inch tip.

**2.9 Gel electrophoresis, protein quantification**

phenol red, 1.25% 2-mercaptoethanol, pH 8.5.

Tris base, 0.1% SDS, 1 mM EDTA, pH 7.2.


*Growing and Handling of Bacterial Cultures within a Shared Core Facility for Integrated… DOI: http://dx.doi.org/10.5772/intechopen.81932*
