Total Collaborative Study Protocol_Solus One Salmonella v1 1

room temperature (15–25°C). Determine the number of wells required for the test. Take the required 

number of strips from the pouch and fit them to the framer provided. Note: Unused strips should be  returned to the pouch and stored at 2–8°C.  ( b ) Prepare the Washing Buffer by adding the contents of a single Washing Buffer Activator  sachet and Washing Buffer bottle (60 mL) to 1440 mL or a single Washing Buffer Activator sachet and 

Washing Buffer bottle (10 mL) to 240 mL of deionized water to prepare the washing buffer at assay 

concentration. Mix until the activator has fully dissolved. 

( c )  Leave the first well in the strip empty to serve as a “blank” for measuring the absorbance of 

the substrate. 

( d ) Aliquot 0.1 mL of the Negative Control in the second well and 0.1 mL of the Positive Control in 

the third well. 

( e ) Add 0.1 mL of each heated sample to each consecutive well in the strip. If there are wells left  over at the end of a test strip the Positive or Negative Controls may be repeated.  ( f )  Incubate the plate (containing the strips) at 37 ± 1°C for 30 ± 5 min.  ( g )  Following incubation, aspirate the contents of the wells, removing as much of the liquid as 

possible. Wash the wells 5–7 times with Washing Buffer ensuring completed filling and emptying of the 

wells through each wash cycle. Note: The washing technique is critical to assay performance; hence it is 

recommended to use a microplate washer.   

( h ) After completion of the washing steps, add 0.1 mL of Conjugate to each well, except for the  blank. Upon completion of the addition of the Conjugate, incubate the plate (containing the strips) at 37 

± 1°C for 30 ± 5 min. 

( i ) 

Repeat the aspiration and wash cycles. 

( j )  After completion of the second aspiration and washing steps, add 0.1 mL of TMB Substrate to

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