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Landscape of Interacting Microbes - DNA Extraction Protocols

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DNA quantification with Qubit™ Flex

Product information: https://www.thermofisher.com/order/catalog/product/Q33327

Protocol written by Shira Jaye Anteby-Maleh

1. Set up and label Qubit™ strip tubes for standard 1 (no DNA), standard 2 (known concentration of DNA), and sample numbers
2. Click “Calculator” on Qubit™ Flex fluorometer -> select reagent -> select 8 wells, # of samples (change according to number of samples being quantified), and 2 standards – this calculates the volume of working solution to pipette into reservoir using 5ml pipette (1ml = 1,000ul)
3. Using a multichannel pipette, add 190ul of working solution from reservoir to each strip tube for standard 1 and standard 2, and add 198ul to each strip tube used for DNA samples
4. Pipette 10ul (using P10) of standard 1 to each reaction tube, and repeat for standard 2
5. Pipette 2ul (using P10) of DNA from each sample to each numbered reaction tube
6. Vortex to mix for 2 sec and spin for 2 sec
7. On the Qubit™ Flex fluorometer, select ds HS DNA, and place each strip tube into the fluorometer, starting with standard 1, then standard 2, and then each numbered strip tube (select read standards and run samples) – select add samples each time a strip is inserted and unselect any unused wells