- 3M NaOAc (5.2)
- NaOAc.3H2O 40.81 g
- DD-H2O 80 ml
- Adjust pH to 5.2 with glacial HOAc, adjust volume to 100 ml with DD-H2O, and autoclave to sterilize.
- Gel-Loading Buffer (6x)
- Bromophenol Blue .025 g
- Xylene Cyanole .025 g
- Ficoll 400 1.5 g
- DD-H2O to 10 ml
- May be stored at room temp or at 4 degrees.
- 1M Tris (per 500 ml)
- Tris Base 60.55 g
- DD-H2O 400 ml
- Dissolve and adjust to the desired pH by adding concentrated HCl.
- Adjust volume to 500 ml with DD-H2O.
- Autoclave to sterilize.
- 0.5M EDTA (8.0) (per liter)
- EDTA.2H2O(disodium salt) 186.1 g
- DD-H2O 800 ml
- Stir vigorously and adjust pH to 8.0 with concentrated NaOH in order to dissolve the EDTA salt.
- Adjust the volume to 1 liter and autoclave to sterilize.
- 20% SDS (per 500 ml)
- In a fume hood, carefully add 1 100 g bottle of SDS to 450 ml DD-H2O.
- Heat to assist dissolution.
- Adjust pH to 7.2 using concentrated HCl.
- Adjust volume to 500 ml with DD-H2O
- sterilize by filtration.
- Ethidium Bromide (10 mg/ml) (per 100 ml)
- In an amber bottle, add 1 g of EtBr to 100 ml of DD-H2O.
- Stir for several hours and store at 4 degrees.
- USE GLOVES
- CaCl2 Solution (per 500 ml)-for preparing competent cells
- CaCl2.2H2O 4.41 g
- Glycerol 75 ml
- 1M Tris 7.5 5 ml
- DD-H2O 400 ml
- Adjust volume to 500 ml with DD-H2O and filter sterilize. Store at 4 degrees.
| - TAE(50x) (per liter)
- Tris Base 242 g
- Glacial HOAc 57.1 ml
- 0.5M EDTA 8.0 100 ml
- Add DD-H2O to 500 ml and autoclave to sterilize.
- GTE (per 100 ml)
- Glucose 0.90 g
- 0.5M EDTA 2 ml
- 1M Tris 8.0 2.5 ml
- Add DD-H2O to 100 ml, filter sterilize and store at 4 degrees.
- TBE(20x) (per liter)
- Tris Base 216 g
- Boric Acid 110 g
- 0.5M EDTA 8.0 80 ml
- Add DD-H2O to 500 ml and autoclave to sterilize.
- KOAc Solution (per 100 ml)
- 5M KOAc 60 ml
- Glacial HOAc 11.5 ml
- DD-H2O 28.5 ml
- Filter sterilize and store at 4 degrees.
- TE(10x) (per 500 ml)
- 1M Tris 8.0 50 ml
- 0.5M EDTA 8.0 5 ml
- DD-H2O 445 ml
- Autoclave to sterilize.
- Prehybridization Solution (per 500 ml)
- NaCl 29.22 g
- Ficoll 400 1 g
- PVP 360 1 g
- BSA 1 g
- NaPyrophosphate 0.5 g
- 1M Tris 7.4 25 ml
- Formamide 250 ml
- Combine all ingredients in a 1 liter flask with stirring.
- Add DD-H2O to just under 500 ml.
- Stir for several hours or overnight
- then add: 2.5 ml 20% SDS
- Before use, heat sonicated sperm DNA to 95 degrees for minutes and add to Prehyb.
- 5M NaCl (per liter)
- 292.2 g NaCl
- 800 ml DD-H2O
- Dissolve, adjust volume to1 liter, and autoclave to sterilize.
- 5M NH4OAc (per 100 ml)
- NH4OAc 38.5 g
- DD-H2O 80 ml
- Adjust volume to 100 ml with DD-H2O and sterilize by filtration.
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