Oct 14, 2020
Banana sensor sfGFP test
This protocol is a draft, published without a DOI.
- 1Chung Shan Medical University
Protocol Citation: Hung Liang Pai 2020. Banana sensor sfGFP test. protocols.io https://protocols.io/view/banana-sensor-sfgfp-test-bh66j9he
Manuscript citation:
https://www.protocols.io/view/toehold-testing-pure-system-protein-production-73ehqje
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: July 02, 2020
Last Modified: October 14, 2020
Protocol Integer ID: 38846
Abstract
The goal of this protocol is to ensure that our lab technique and environmental circumstarnces are compatible to this expiriment. Based on iGEM team EPFL 2019, we try to define a concentration of Toehold switch and trigger DNA sequence which has a high protein expression(sfGFP in this case) but low or no leakage at same time.
Guidelines
All the materials are sensible to temperature, as a result, keep them on ice. In order to keep all the wells starting at the same time, drop different solution and reagent to their own corner seperately (follow the picture).
Preparation
Preparation
Sterilize the bench, and put on a labmat
Thaw the reagents On ice
Set the microplate (384 well plate) reader machine
Protocols
Protocols
Spin down or gently shake the tube before adding (especially solution A and DNA plasmid)
Separate the droplets in the different corners of the well (as the picture below)
This picture is from iGEM EPFL
Seal the plate with microseal On ice
Centrifuge 4000 rpm ,00:01:00 , 4 °C
Place the plate in the readers, make sure the setting is correct, and start the measurement
After 02:00:00 of measurement, save the data, and turn off all the machines