Buffers used in dna overstretching and unzipping experiments

Buffers used in DNA-overstretching and unzipping experiments

• Popping Buffer

• PBS

We primarily use popping buffer and PBS for DNA-unzipping and overstretching experiments

Popping Buffer

Chemicals used in popping buffer:• EDTA:

• Sodium Phosphate• Monobasic:

• Dibasic

• NaCl

• Tween20:

• Water H2O:

• D2O

Quantities and measurements

CalculationsLabView

Weight & Measure

Store

PBS

Chemicals used in PBS: PBS and H2O or D2O

• Dissolve 1 table in 200mL of H2O or D2O

BGBBlotting 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.

Filter

Anti-Dig

Microspheres

DNA