Cloning of Genomic Fragments

1. Lambda Zapll Cloning and Packaging Kit (Stratagene, La Jolla, CA, USA): RecA-Escherichia coli host strain XL1-Blue MRF'; ExAssist interference-resistant helper phage; SOLR strain; Packaging extracts.

2. 10X Ligase Buffer: 500 mM Tris-HCl, pH 7.5, 70 mM MgCl2, 10 mM dithiothreitol (DTT) plus.

4. T4 DNA Ligase (Epicentre).

5. SM Buffer (l L): 5.8 g NaCl, 2.0 g MgSO4 ■ 7H2O, 50 mL 1 M Tris-HCl, pH 7.5, 2 mL 5% (w/v) gelatin, add H2O to 1 L and autoclave.

6. Chloroform.

7. LB Plates (For 1 L): 10 g NaCl, 10 g tryptone, 5 g yeast extract, 20 g agar adjust to pH 7.0.

8. LB Media (For 1 L): 10g NaCl, 10 g tryptone, 5 g yeast extract, adjust to pH 7.0.

9. LB-Tetracycline Plates: LB plates as above with tetracycline 12.5 ^g/mL.

10. LB-MgSO4-Maltose Media: LB media as above with 10 mM MgSO4 and 0.2% maltose.

11. Top Agar: Prepare 1 L of LB broth and add 0.7% [w/v] agarose, autoclave.

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