AOAC-RI ERP Book MICRO.pdf

Page

3) Prepare the reconstituted assay by adding 1040 µL(96 sample kit) or 130 µl (24

sample kit) of the Multiplex PCR master mix to a vial of

mericon

Assay and mix

by pipetting up and down 2-3 times. Prepare a positive control by re-suspending

the lyophilized Positive Control DNA in 200µL of QuantiTectNucleic Acid Dilution

Buffer and mix. Prepare the appropriate number of strip tubes by transferring

10µL of the reconstituted assay to the reaction tubes. Pipette 10µL of the

extracted sample DNA into each sample strip tube and mix by pipetting up and

down 2-3 times. Prepare the positive control tube by pipetting 10µL of the

Positive Control DNA into a strip tube containing 10 µL of reconstituted assay.

Prepare the negative control by pipetting 10 µL of RNase-free water into a strip

tube containing 10 µL of thereconstituted assay. Seal the strip tubes with the

strip tube caps. Load the samples into the Rotor-Gene-Q and initiate the analysis

immediately.

(b) Automated QIAsymphonyDNA Extraction Method

1) Ensure the “Waste” drawer is prepared properly, and perform an inventory scan

of the “Waste” drawer, including the tip chute and liquid waste. Replace the tip

disposal bag, if necessary.

2) Load an elution microtube rack (EMTR) into the “Eluate” drawer and perform an

inventory scan of the “Eluate” drawer.

3) Load the

QIAsymphonymericon

bacteria kit and consumables into the “Reagents

and Consumables” drawer.Select the “R+C” button on the touchscreen to open

the screen that shows the consumables status (“Consumables/8-

RodCovers/Tubes/ Filter-Tips/Reagent Cartridges”).Select the “Scan Bottle”

button to scan the bar code of the TopElute bottle with the handheld bar code

scanner. Select the “OK” button.

4) Perform an inventory scan of the “Reagents andConsumables” drawer.

5) Transfer 500 µL of the enriched sample into a 2 mL screw cap tube.Place the

samples into the appropriate tube carrier and load them into the “Sample”

drawer.

6) Using the touchscreen, enter the required information for each batch of samples

to be processed.

7) Choose elution volumes according to Table 1.

Table 1: Matrix Manual Dilution and Automated Elution Volumes

Matrix

Manual

mericon

Method

DNA Dilution

Automated

QIAsymphony

Method

DNA Elution Volume

Fresh Spinach

1:10

400 µL

OMAMAN-36 E/ AOAC PTM Report

ERP Use Only

January 2017

AOAC Research Instit te

Expert Review Pa el Use Only