Screening for lacZ Transgene Expression

1. Positively charged nylon membrane (Boehringer Mannheim 1417240).

4. Buffer 3 (2X SSC): This should be prepared from a 20X SSC (3 M NaCl/0.3 M Na-citrate2H2O) stock solution adjusted to pH 7.0 with 1 M HCl.

5. Stratalinker UV crosslinker (Stratagene, La Jolla, CA).

7. Pre-hybridization buffer: 5X SSC, 0.1% N-lauroylsarcosine, 0.02% SDS, 3% blocking reagent (Boehringer Mannheim 1096176).

11. Anti-digoxigenin (DIG) antibody (Boehringer Mannheim 1093274) diluted 1:10,000 in buffer 6 containing 3% blocking reagent.

13. Buffer 7: 0.1 M Tris-HCl, pH 9.5, 0.1 M NaCl, 0.05 M MgCl2. Prepare fresh on day of use.

14. CSPD (25 mM; Di-sodium, 3-(-4 metho-xyspiro[1,2-dioxetene-3,2'-{5' chloroj tricyclo {3.3.1.1 j decan]-4-yl)phenyl phosphate; Boehringer Mannheim 1655884) diluted 1: 100 in buffer 7 before use.

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