1. Equipment and software.

a. Micropipets.

c. Incubator-shakers (New Brunswick Scientific Series 25D).

d. Water baths.

e. Hot/stir plate.

f. Heat block.

g. Multitube vortexer (VWR 58816-115).

h. Benchtop vortexer.

i. Refrigerated superspeed centrifuge (Sorvall RC-5B).

j. Swinging-bucket rotor with tube holders (Sorvall SH3000).

k. Microcentrifuge (Eppendorf 5415C).

l. Spectrophotometer.

m. Thermocycler with an ALD-1234 dual 30/48 position block alpha unit with heated lids (MJ Research PTC-200).

o. Vac-Man vacuum manifold (Promega).

p. Sequencing gel box and mini-gel boxes (Owl Model S3S).

r. UV transilluminator.

s. Light imaging system and software (Alpha Innotech Alphalmager 950).

t. Phosphor screen (Molecular Dynamics).

u. Phosphorimager (Molecular Dynamics Storm 860).

v. ImageQuant software.

w. Comparable equipment/software may be substituted. 2. Sterile consumables.

a. 1250-^L Micropipet tips (VWR 53508-924).

b. 200-^L Micropipet tips (VWR 30128-376).

c. 50-mL Conical centrifuge tubes (VWR 21008-242).

d. 15-mL Conical centrifuge tubes (VWR 21008-216).

e. 1.7-mL Microcentrifuge tubes (Costar 3620).

f. 1.5-mL Screw-cap microcentrifuge tubes (VWR 20170-215).

g. 0.65-mL microcentrifuge tubes (Costar 3208).

h. 0.5-mL Thin-wall PCR tubes (MJ Research TBI-0521).

i. 1.2-mL Cryogenic storage vials (Corning 430658). j. 10-mL Pipets (BD Falcon 357551).

k. 5-mL Pipets (BD Falcon 357543).

l. 0.45-^m Bottle-top filters (Nalgene 295-3345 or Corning 430625).

m. Wood applicator sticks (Puritan 807).

n. 1-mL Slip-tip syringe barrels (BD 309602).

o. 0.45-^m syringe filters (Whatman 6780-2504).

p. Nonsterile disposable cuvetes (VWR 58017-847).

q. Comparable consumables may be substituted.

Unless otherwise specified, solutions are prepared using water that has been purified to a resistance of 18.2 Mfl-cm (dH2O).

Caution: A number of hazardous compounds are used in these protocols, including but not limited to 17^-estradiol, chloroform, phenol, sodium metabisulfite, sodium hydroxide, hydroquinone, [y 32P]ATP, unpolymerized acrylamide, and TEMED. Personnel should be trained in the use of the materials listed here and should take appropriate precautions as set forth by the manufacturer and institutional policy.

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