Analysis of Differential Clones

1. Random oligonucleotide labeling solutions (see Note 7).

2. Deionized formamide (Gibco-BRL).

3. 37% Formaldehyde (Sigma-Aldrich Chemicals, St. Louis, MO).

4. 10X MOPS buffer; 0.4 M MOPS pH 7.0, 0.1 M sodium acetate,10 mM EDTA.

5. 6X gel loading buffer: 25% (w/v) Ficoll 400, 0.25% (w/v) bromophenol blue, 0.25% (w/v) xylene cyanol, 1 mM EDTA.

8. MAGNA NT nylon hybridization membranes (Micron Separations, Inc., Westboro, MA) or equivalent.

10. UV crosslinker or 80°C oven.

11. 50X Denhardt's solution: 1% (w/v) bovine serum albumin, 1% (w/v) polyvinyl pirrolidone, 1% (w/v) ficoll 400. Filter-sterilize and store in aliquots at -20°C. Thaw at room temperature when needed. Can be temporarily stored at 4°C.

13. 20% Sodium dodecyl sulfate (SDS).

14. TA Cloning kit (Invitrogen) (see Note 9).

15. DNA sequencing kit (USB, Cleveland, OH).

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