| 5M NaCl | |
| NaCl | 292 g |
| milliQ dH2O | up to 1000 mL |
| TOTAL | 1L |
| 2M KCl | |
| KCl | 149.1 g |
| milliQ dH2O | up to 1000 mL |
| TOTAL | 1L |
| 0.5 M EDTA pH 8.0 | |
| EDTA disodium dihydride | 186.1 g |
| Stir in 800 mL dH20 and add ~20g NaOH pellets until the pH reaches 8 and the EDTA fully dissolves. Will dissolve as the pH increases. | |
| milliQ dH2O | up to 1000 mL |
| TOTAL | 1L |
| 2X Laemmli sample buffer (store at RT) (add BME to 10% before use) | |
| 20% SDS | 10 mL |
| glycerol | 10 mL |
| 1M TRIS-HCl | 6 mL |
| milliQ dH2O | 24 mL |
| Bromophenol blue | Tiny amount (tip of spatula) |
| TOTAL | 50 mL |
| 6X DNA Loading Buffer | |
| Bromophenol blue | 100 mg |
| xylene cyanol FF | 100 mg |
| Ficoll 400 | 6 g |
| milliQ dH2O | up to 40 mL |
| TOTAL | 40 mL |
| TE (Tris-EDTA) | |
| 1M TRIS pH 8 | 500 uL |
| 0.5M EDTA | 100 uL |
| milliQ dH2O | up to 50 mL |
| TOTAL | 50 mL |
| RIPA | |
| NaCl | 8.76 g |
| 1M TRIS pH 7.2 | 10 mL |
| 20% SDS | 5 mL |
| sodium deoxycholate | 10 g |
| 0.5M EDTA | 10 mL |
| milliQ dH2O | up to 1L |
| TOTAL | 1L |
| Coomassie Stain | |
| MeOH | 100 mL |
| Acetic acid | 20 mL |
| Brilliant Blue R-250 | 100 mg |
| dH2O | 80 mL |
| TOTAL | 200 mL |
| Coomassie Destain |
||
| MeOH | 80 mL | 400 mL |
| Acetic acid | 20 mL | 100 mL |
| dH2O | 100 mL | 500 mL |
| TOTAL | 200 mL | 1000 mL |
| 10X PBS 4L | ||
| NaCl | 80 g | 320 g |
| KCl | 2 g | 8 g |
| Na2HPO4 | 14.4 g | 57.6 g |
| KH2PO4 | 2.4 g | 9.6 g |
| dH20 | 1 L | 4 L |
| pH should be around 6.6 which will dilute to ~7.4 when at 1X | ||
| TOTAL | 1L | 4 L |
| 50x TAE | |
| TRIS Base | 242 g |
| Acetic acid | 57.1 mL |
| 0.5M EDTA pH 8.0 | 100 mL |
| milliQ dH20 | Until 1000 mL mark |
| TOTAL | 1000 mL |
| 10x TBE |
||
| TRIS Base | 108 g | 1080 g |
| Boric acid | 55 g | 550 g |
| 0.5M EDTA pH 8.0 | 40 mL | 400 mL |
| milliQ dH20 | Until 1L mark | Until 10L mark |
| TOTAL | 1L | 10 L |
| 10X SDS-PAGE Running Buffer | |
| TRIS Base | 600 g |
| Glycine | 1440 g |
| SDS | 100 g |
| milliQ dH20 | Until 10L mark |
| TOTAL | 10 L |
| 1X TBST | |
| TRIS base | 12.2 g |
| NaCl | 87.65 g |
| Tween-20 | 5 mL |
| Add some dH20, but leave room for NaOH solution. Use NaOH to get pH to 8.0 | |
| milliQ dH2O | up to 1L |
| TOTAL | 10L |
| Luria Broth (LB) |
|||
| Bacto-tryptone | 10 g | 50 g | 100 g |
| Bacto-yeast extract | 5 g | 25 g | 50 g |
| NaCl | 10 g | 50 g | 100 g |
| milliQ dH20 | Until 1L mark | Until 5 L mark | Until 10 L mark |
| TOTAL | 1 L | 5 L | 10 L |
| Luria Broth (LB) Agar Plates |
||
| Bacto-tryptone | 10 g | 20 g |
| Bacto-yeast extract | 5 g | 10 g |
| NaCl | 10 g | 20 g |
| Agar | 20 g | 40 g |
| milliQ dH20 | Until 1L mark | Until 2 L mark |
| Autoclave the solution. If adding antibiotics wait until it cools slightly (~55C) before adding the antibiotic. Mix and pour into plates (~25mL/plate). | ||
| TOTAL | 1 L (~40 plates) | 2 L (~80 plates) |
| Terrific Broth |
|||||||||||
| yeast extract | 24 g | ||||||||||
| Tryptone | 20 g | ||||||||||
| Glycerol | 4 mL | ||||||||||
| milliQ dH20 | 900 mL | ||||||||||
|
Autoclave the solution. Seperately prepare:
Add 100 mL of the sterile buffer solution to the 900 mL of TB. |
|||||||||||
| TOTAL | 1 L | ||||||||||
For E. Coli:
| Antibiotic | 1000X stock | Working Concentration |
|---|---|---|
| Ampicillin | 100 mg/mL | 100 µg/mL |
| Chloramphenicol | 25 mg/mL (dissolve in EtOH) |
25 µg/mL |
| Hygromycin B | 200 mg/mL | 200 µg/mL |
| Kanamycin | 50 mg/mL | 50 µg/mL |
| Spectinomycin | 50 mg/mL | 50 µg/mL |
| Tetracycline | 10 mg/mL | 10 µg/mL |