One-pot lyophilized SHERLOCK assay

Prepare lyophilized one pot SHERLOCK pellets:

Reagent manufacturers and further instructions are listed in the methods section in the Science Advances publication

1 - Prepare one-pot SHERLOCK solution using the following components into a 0.2mL PCR tube and mix well: 

    1 - TwistAmp Basic pellet (Twist Dx)

    2 - 0.24uL of forward and reverse RPA primers each from a 100uM stock concentration 

    3 - 30uL of rehydration buffer (33mM HEPES pH 6.8, 100mM NaCl, 8.3% PEG) 

    4 - 2.5uL of Protoscript reverse transcriptase from a 100uM stock concentration

    5 - 0.25uL of Ambion RNase H from a 10U/uL stock concentration 

    6 - 0.5uL of ssDNA probe 

    7 - 0.2uL of EnGen Lba Cas12a from a 50uM stock concentration 

    8 - 0.2uL of gRNA solution from a 100uM stock concentration 

    9 - 5uL of 10x NEB 2.1 buffer

2 - Punch a hole into the 0.2mL PCR tube cap using an 18 Gauge needle 

3 - Snap freeze the above PCR tube in liquid nitrogen 

4 -  Quickly re-open snap-frozen PCR tube and add 2.5uL of Magnesium acetate from a 280mM stock concentration 

5 - Snap freeze the PCR tube again 

6 - Lyophilize the snap-frozen PCR tube for 6 hrs 

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