Preparation of Lyophilized CRISPR-Cas13a Premixed Detection Reagents for Nucleic Acid Biomarkers Detection

To begin, prepare the CRISPR-Cas13a master mix reaction using the components shown here. After thoroughly mixing the contents, spin down the master mix tube. Aliquot 17 microliters of the master mix into ten 1.5 milliliter tubes placed on ice.

Place the tubes in a rack, submerged in liquid nitrogen, and allow them to remain submerged for five minutes. At the freeze dryer check collector and shelf temperatures, then place the tubes in the freeze dryer. Select the program and press start.

After 30 minutes of the secondary drying step, stop the program. To release the pressure open the vacuum release valve and remove the rack from the shelf. Cap the tubes immediately before storing the lyophilized reactions at minus 20 degrees Celsius.