Storage
@Yujeong
Lyophilization
- The reagents of the reaction were lyophilized to three tubes; the RAA mix (in our case, this will be RPA mix), the RAA buffer mix, and Cas mix.
- RAA mix: supplied from the RAA kit
- RAA buffer mix: primers, buffer for the solution for a 50ul reaction
- Cas mix: ssDNA reporter, cas reagents
- Lyophilization Procedure:
- RAA buffer mix and Cas mix were pre-frozen at -80°C for 1-3h
- Then, freeze-dried at -50°C overnight
- Nucleic acid detection Procedure via DETECTR:
- Dissolve the RAA mix into a 50μL reaction, containing dsDNA target
- Transfer the solution to the RAA buffer mix
- Incubate at 37°C for 30 minutes
- Add 2-20μL of the solution to the Cas mix
- Incubate at 37°C for 30 minutes
- They remained the same sensitivity when stored at 4°C for at least 7 days.
- Lyophilized reagents can withstand long-term transportation and storage. Hence, it is good for field-based applications.
- "The lyophilization significantly simplifies the operational process and improves the detection stability" (Bai, 2019)
- "The future development of portable microfluidic-based cartridges and lyophilized reagents to run the assay could enable point-of-care testing outside of the clinical diagnostic laboratory, such as airports, local emergency departments and clinics and other locations." Nature (Broughton, 2020)
- The reagents of the reaction were lyophilized to three tubes; the RAA mix (in our case, this will be RPA mix), the RAA buffer mix, and Cas mix.
https://link.springer.com/article/10.1007/s10544-010-9423-4#Sec2
- Liquid/ dry reagents are stored inside the microfluidic chips