Performing Tests - Quench–Gone Aqueous
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QGA uses a single test to isolate Cellular ATP (ATP from living cells) in a water sample. This eliminates ATP from cells that have died and therefore provides a true measure of biological activity.
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cATP Analyses using QGA consists of the following steps:
Mix your sample to ensure homogenity. Remove the plunger from a 60mL syringe and attach a filter. Pour the appropriate volume of sample from 2.1 into the barrel of the syringe.
TIP: Alternatively, the syringe tip can be immersed into the sample and drawn into the barrel before attaching the filter, rather than pouring the sample into the barrel after attaching the filter. Be sure the syringe tip is clean if this mechanism is used.
Slowly push the entire sample volume through the filter and into a waste receptacle. Push the plunger far enough to filter the sample and stop to ensure that the filter remains wet. Detach the filter and remove the plunger.
NOTE: If the full volume of sample could not be filtered, record the actual volume processed.
TIP: If increased sensitivity is desired, filter additional sample by repeating 2.2 and 2.3 using the same syringe and filter.
Re-attach the filter to the syringe barrel. Use the micropipettor to add 1mL of UltraLyse 7 to the barrel. Pass the UltraLyse 7 through the filter to dryness and collect in an unused 17x100mm test tube. This is called the Extraction Tube.
NOTE: At this point, the contents of the Extraction Tube can be capped and stored refrigerated between 2-8 oC for up to 1 week prior to 2.5 and 2.6.
Add 9mL of UltraLute into the Extraction Tube. Cap and invert three times to mix. This is now called the Diluted Extraction Tube.
NOTE: At this point, the contents of the Diluted Extraction Tube are stable at room temperature for up to 4 hours.
Using the micropipettor, transfer 100µL of the Diluted Extraction Tube contents to a new 12x55mm test tube (the Assay Tube), add 2 drops (100µL) of Luminase, swirl gently five times, and insert into the Luminometer. Record RLUcATP for use in the final calculations.
NOTE: If RLUcATP < 10 on a Kikkoman Lumitester C-100 or C-110, you are below the low–detection limit. Report cATP (pg ATP/mL) = 0 in the final calculations, or select a larger volume in 2.1 and repeat the analysis.
TIP: If “Scale Over” is returned, repeat the analysis using a smaller sample volume in 2.1.
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UltraCheck1 Standard
For each set of tests, an UltraCheck1 Standard must also be performed. This is used as a calibration to convert RLU values obtained from ATP assays into actual ATP concentrations:
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