Acrylamide Solutions:
- -Filter all solutions with .45µm
| Percentage: |
6% |
8% |
14% |
20% |
23% |
| 5X TBE: |
50ml |
50ml |
50ml |
50ml |
50ml |
| Urea: |
210.21g |
210.21g |
210.21g |
210.21g |
210.21g |
| 40% Stock Acrylamide.: |
75ml |
100ml |
175ml |
250ml |
288ml |
| Total Volume: |
500ml |
500ml |
500ml |
500ml |
500ml |
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0.5M EDTA:
- 10g NaOH
- 400ml ddH2O
- add 93g Na2EDTA while stirring
- Increase Vol to 500ml with ddH2O
- Check pH to 8.0, aliquot into 50ml conicals
1X TE:
- 0.5ml 2M Tris (8.0) - 10mM final
- 0.2ml 0.5M EDTA - 1mM final
- ddH20 to final volume of 100ml. TE Low has 0.1mM EDTA.
5X TBE:
- 54g Tris Base
- 25.5g Boric Acid
- 20ml 0.5M EDTA (8.0)
- ddH2O to final volume of 1L. (pH 8.3)
50X TAE:
- 242g Tris Base
- 57.1ml Glacial Acetic Acid
- 100ml 0.5M EDTA (pH 8.0)
- water to 1L
FLB: (Formamide Loading Buffer)
- 4.75ml 95% Formamide
- 150µL 0.5M EDTA
- 50µL 2.5% Bromophenol Blue
- 50µL 2.5% X.C.
- Make 100µL aliquots, filter thru large syringe, 0.2µm.
Reverse Transcriptase (RT) Buffer:
| 25mM KCl |
83µL 3M KCl |
| 50mM Tris (7.5) |
250µL 2M Tris |
| 10mM DTT |
100µL 1.0M DTT |
| 3.5mM MgCl2 |
35µL MgCl2 |
| 0.5mM all 4 dNTPs |
50µL each dNTP @ 100mM |
| 100µg/mL BSA |
50µL 20mg/mL BSA |
| ddH2O |
to 10mL |
- Filter, make ~600µL aliquots