Update on NC detection method
What has been accomplished thus far:
- Able to detect BSA on NC membrane with the use of Ponceau S stain
- With PDMS array on NC, no BSA leaks through the area covered by PDMS
- Initial trial done with 10:1 ratio of elastomer base: curing agent. Later 3:1 and 5:1 ratios done. 3:1 is too sticky even after heating in the oven for 2+ hours
- two ways to stamp PDMS onto NC, array created using toothpicks and sepcons and negative stamp used with sepcons on NC and PDMS poured around. The negative stamp works the best and allows for the least amount of PDMS spreading but is more difficult to work with, since the sepcons can shift on the NC in transport
Where to go from here:
- W677 w and w/o pinholes to pass BSA onto the NC membrane and stain with Ponceau S
- first make sure the membranes will fit on the PDMS array- alter array if needed
- use 1X PBS on the backside of the membrane and 1 mg/mL BSA on the membrane
- Later set up 3 membranes, each with BSA, Myosin and a BSA + Myosin mixture. If the set-up works, when stained BSA and BSA + Myosin will be show a pink detection.
- Electrophoretic array
- Set up a pressurized model
- Once the set-up works with BSA and Myosin, use Harold Smith’s proteins to test the detection method.
- Create a new mask design for the PDMS array.