Buffers used in dna overstretching and unzipping experiments
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The document outlines the preparation of buffers for DNA overstretching and unzipping experiments, primarily focusing on popping buffer and PBS. It details the chemicals used in each buffer and provides specific instructions for preparing a blotting grade blocker solution in the popping buffer. Storage instructions and shelf life of the prepared solutions are also mentioned.
Buffers used in dna overstretching and unzipping experiments
- 1. Buffers used in DNA-overstretching and unzipping experiments
- 2. We primarily use popping buffer and PBS for DNA-unzipping and overstretching experiments • Popping Buffer • PBS
- 3. Popping Buffer Chemicals used in popping buffer: • EDTA:
- 4. • Sodium Phosphate • Monobasic:
- 5. • Dibasic
- 6. • NaCl
- 7. • Tween20:
- 8. • Water H2O:
- 9. • D2O
- 12. Weight & Measure
- 13. Store
- 14. PBS Chemicals used in PBS: PBS and H2O or D2O • Dissolve 1 table in 200mL of H2O or D2O
- 15. BGB Blotting Grade Blocker To make 5mg/ml BGB solution in popping buffer: •First wiight out 15mg of BGB poweder •Add 1.5 ml of 1x pop and mix by vortexing •Run through .2μM syringe filter using 3ml syringe. •Now store 5mg/ml BGB in 1x pop buffer (minus amount of protein that stuck to filter) at 3C. This should be good for few weeks.
- 16. Filter
- 17. Anti-Dig
- 18. Microspheres
- 19. DNA