Outer solution

Scope: how-to cookbook guide to take into lab and make outer solutions for future use in upstream protocols.

Glucose stocks for outer solution

• [ ] (Simple) Outer solution: 800mM glucose
  • [ ] Add 1.44 g glucose and roughly 9.2mL MilliQ water to a final volume of 10mL in a graduated cylinder.
  • [ ] Transfer to 15-mL Falcon tube and vortex until dissolved.
• [ ] (Stock) Outer solution: 3 M glucose
  • [ ] Combine 5.40 g glucose (MW = 180.16 g/mol) and roughly 5.0 mL MilliQ water in a 15 mL tube.
  • [ ] Heat and mix vigorously as necessary to dissolve material completely. (consider adding a picture showing dissolution).
  • [ ] Add additional MilliQ water to achieve a final volume of 10 mL.
  • [ ] Filter sterilize and store at -20 C.

Sucrose stocks for inner solution

• [ ] (Simple) Inner solution: 800mM sucrose
  • [ ] Add 2.81 g sucrose and roughly 8.0mL MilliQ water to a final volume of 10mL in a graduated cylinder.
  • [ ] Transfer to 15-mL Falcon tube and vortex until dissolved.
• [ ] (Stock) Inner solution: 2 M sucrose
  • [ ] Combine 10.27 g sucrose (MW = 342.3 g/mol) and roughly 5.0 mL MilliQ water in a 15 mL tube.
  • [ ] Heat and mix vigorously as necessary to dissolve material completely.
  • [ ] Add additional MilliQ water to achieve a final volume of 15 mL.
  • [ ] Filter sterilize and store at -20 C.

HPTS stock for simple trouble shooting

• [ ] Make 1mL of 4mM HPTS. Add 2.1 mg HPTS and 1mL MilliQ water.

NOTES: Combine sucrose and HPTS solutions at a 19:1 ratio as needed.

NOTES: brightness depends significantly on pH

Reference table