FAQ

KASP genotyping reactions consist of three components: universal KASP Master mix, SNP-specific KASP Assay mix, and template DNA. The reactions can be prepared using either wet DNA (Table 1) or dried down DNA (Table 2), depending on your preferred laboratory workflow.

Table 1: Wet DNA reaction assembly

Table 2: Dry DNA reaction assembly 

The volumes outlined in the tables above give the exact ratios of the reaction components that should be used when preparing KASP reactions. It is expected that reactions will not be assembled individually, but that larger volumes (sufficient for all planned reactions plus an additional percentage to account for pipetting error) will be prepared prior to dispensing into the reaction plate.

Please note that it is possible to round up the volumes of DNA / water used in the reactions if preferred (e.g. from 2.43 µL to 2.5 µL for 5 µL reactions). The below example details how to prepare reactions in this way, if setting up a full 96-well plate of reactions using wet DNA.

1. Dispense 5 µL of template DNA at the appropriate concentration into each well of the 96-well plate. Please note that at least two wells of the plate should contain water instead of DNA – these wells will act as the no template controls (NTCs).
2. Combine KASP Master mix and KASP Assay to create KASP genotyping mix. The table below details how to prepare sufficient KASP genotyping mix for 96 reactions plus 10% excess.

   	KASP genotyping reaction assembly (dry DNA)
   Component	5 µL reaction  (384-well plate)	10 µL reaction (96-well plate)	96 x 10 µL reactions plus 10% excess
   KASP Master mix (2X)	2.5 µL	5 µL	528 µL
   KASP Assay mix (72X)	0.07 µL	0.14 µL	14.78 µL
   Total volume	5 µL	10 µL	542.78 µL

3. Pipette 5 µL of KASP genotyping mix (see table above) into each well of the reaction plate. Combined with the template DNA, this gives a final reaction volume of 10 µL per well.