1. Lysis Buffer
| Component | Concentration | Amount |
|---|---|---|
| KCl | 1M | 50µl |
| Tris (pH 8.3) | 1M | 10µl |
| MgCl2 | 1M | 2.5µl |
| NP-40 | - | 4.5µl |
| dH20 | - | 928.5µl |
2. Agar pads-4% (for microscopy)
3. DEPC-dH20
4. Ethidium Bromide
5. L Broth
| Component | Amount |
|---|---|
| Tryptone | 5g |
| NaCl | 5g |
| Yeast Extract | 2.5g |
| dH20 | 490mL |
| ** Autoclave ** Autoclave ** Autoclave ** | |
6. S Basal Media
| Component | Concentration | Amount |
|---|---|---|
| Di-Potassium phosphate (pH 6) | 0.05 M | 2.43g |
| Potassium dihydrogen phosphate (pH 6) | 0.05 M | 4.98g |
| NaCl | 0.1M | 5.85g |
| Cholesterol (5mg/mL in EtOH) | - | 2mL |
| dH20 | - | 985mL |
| ** Autoclave ** Autoclave ** Autoclave ** | ||
7. SM Media
| Component | Concentration | Amount |
|---|---|---|
| S basal media | - | 1L |
| Potassium Citrate (pH 6) | 1M | 10mL |
| Trace Metals Solution | - | 10mL |
| CaCl2 | 1M | 3mL |
| MgSO4 | 1M | 3mL |
8. 8P plates
| Component | Amount |
|---|---|
| NaCl | 3g |
| Agar | 25g |
| Peptone | 20g |
| dH20 | 970ml |
| ** Autoclave ** Autoclave ** Autoclave ** | |
| Cholesterol (5mg/mL in EtOH) | 1mL |
| CaCl2 (1M) | 1mL |
| MgSO4 (1M) | 1mL |
| 1M Potassium Phosphate (pH 6) | 25mL |
9. M9 Buffer
| Component | Amount |
|---|---|
| KH2PO4 | 3g |
| Na2HPO4 | 6g |
| NaCl | 5g |
| dH20 | 990mL |
| ** Autoclave ** Autoclave ** Autoclave ** | |
| MgSO4 (1M) | 1mL |
10. Proteinase K Buffer
| Component | Final Concentration | Amount |
|---|---|---|
| NaCl (4M) | 50mM | 12.5µl |
| Tris (1M) (pH 7.5) | 50mM | 50µl |
| EDTA (0.5 M) | 5mM | 10µl |
| SDS (10%) | 0.5% | 50µl |
| Proteinase K (20mg/mL) | 200µg/mL | 10µl |
| dH20 | - | 867.5µl |
11. Solution A
| Component | Final Concentration | Amount |
|---|---|---|
| EDTA (0.5M) (pH 8) | 10mM | 20µl |
| SDS (10%) | 0.5% | 50µl |
| dH20 | - | 930µl |
12. Solution B
| Component | Final Concentration | Amount |
|---|---|---|
| EDTA (0.5M) (pH 8) | 10mM | 20µl |
| NaOAc (1M) | 0.1M | 100µl |
| dH20 | - | 880µl |
13. Solution C
| Component | Amount |
|---|---|
| Tris-HCl (pH 7.5) | 440µl |
| NaCl (5M) | 180µl |
14. Proteinase K Solution
| Component | Concentration |
|---|---|
| KCl | 500mM |
| Tris-HCl (pH 8.3) | 10mM |
| MgCl2 | 2.5mM |
| NP-40 | 0.45% |
| Tween-20 | 0.45% |
| Gelatin | 0.01% |
| Proteinase K | 300µg/mL |
15. 5x Loading Buffer
| Component | Amount |
|---|---|
| 10x TBE | 5mL |
| Glycerol | 4mL |
| Bromophenol Blue | 5mg/mL |
| dH20 | 1mL |
16. Solution I
| Component | Final Concentration | Amount |
|---|---|---|
| Glucose | 50mM | 9g |
| Tris-HCl (pH 8.0) | 25mM | 3.9g |
| EDTA (pH 8.0) | 10mM | 3.7g |
17. Solution II
| Component | Final Concentration | Amount |
|---|---|---|
| NaOH (10N) | 0.2 N | 10mL |
| SDS | 1% | 5g |
| dH20 | - | 480mL |
18. Solution III
| Component | Final Concentration | Amount |
|---|---|---|
| Potassium Acetate (5M) | 3M potassium, 5M acetate | 600mL |
| Glacial Acetic acid | - | 115mL |
| dH20 | - | 285mL |
19. 2x Transformation & Storage Solution (TSS)
| Component | Final Concentration | Amount |
|---|---|---|
| Polyethylene Glycol 3350 | 20% w/v | 200g |
| MgCl2 | 100mM | 20.33g |
| ** Autoclave ** Autoclave ** Autoclave ** | ||
| DMSO | 10% v/v | 100mL |