Downloads: Protocols for automated nucleic acid extraction with ASSIST PLUS

Here you can download the complete protocols and reports used in this application and use them on your ASSIST PLUS. The protocol can be customized with the VIALAB software.

Experimental set-up: Program 1

Step-by-step procedure

1. Aliquot the blood samples

Transfer the blood samples from the collection tubes.

Place the blood collection tubes on position C and a 700 μl deep well plate on position B. Then select the ARCIS_SAMPLES program on the VOYAGER electronic pipette by pressing Run. The ASSIST PLUS immediately starts to transfer blood from the collection tubes into the deep well plate.

Tip: Use the Tip Travel feature of the ASSIST PLUS to keep the tip immersion depth constant during aspiration and dispensing, ensuring contamination-free and consistent liquid handling for more reproducible results, no matter how often the application is repeated.

Experimental set-up: Program 2

Step-by-step procedure

1. Reagent distribution

Aliquot Reagent 1 and Reagent 2.

Place two 10 ml multichannel reagent reservoirs into a dual reservoir adapter from INTEGRA on position A, the deep well plate with the preloaded blood samples on position B, and a new 96 well plate on position C. Fill the two 10 ml multichannel reagent reservoirs with Reagent 1 and 2, and change the program on the VOYAGER pipette to ARCIS_KIT. The ASSIST PLUS then starts to aliquot 150 μl of Reagent 1 in to the deep well plate (position B, columns 7-12) followed by the distribution of 20 μl of Reagent 2 in the 96 well plate (position C, columns 1-6).

Tips:

• A pre- and post-dispense step is recommended to increase the accuracy and precision of pipetting. The pre- and post-dispense volume should be between 3 and 5 % of the nominal volume of the pipette.
• Save reagents by using the 10 ml multichannel reagent reservoir with SureFlo anti-sealing array. This allows you to aspirate from the very bottom of the reservoir in order to minimize the dead volume of the reagents.

2. Sample lysis

Add blood samples to Arcis Reagent 1.

Transfer 90 μl of each blood sample to the wells containing Reagent 1. After mixing, there is a one-minute incubation period before the samples are further processed or stored for processing later, according to the manufacturer’s protocol (Arcis Sample Prep Kit).

Tip: Be sure to mix Reagent 1 thoroughly with the blood samples to ensure a homogenous solution.
Note: Nucleic acids are stable for 90 days at room temperature, provided there is no further processing.

3. Sample washing

Wash lysed samples with Arcis Reagent 2.

Continuing this protocol, use the ASSIST PLUS to automatically transfer 5 μl of the lysed blood samples (position B) to Reagent 2 in the new 96 well plate (position C) and mix them thoroughly.

Tip: After addition of Reagent 2, the samples must be used immediately or frozen at -20 ºC. To reduce sample dilution, samples can be added to Reagent 2 at a ratio of 1:3, 1:2 or 1:1.

Remarks

• The PCR reaction can be run immediately or go directly to other downstream techniques. To continue with a PCR reaction, add the appropriate volume to the PCR master mix (e.g. 5 μl per 25 μl of PCR reaction).
• Users who only want to extract DNA from eight samples can place an 8 well PCR strip in a PCR 96 well cooling block (INTEGRA) instead of a 96 well plate.
• To process a higher number of samples consider using the kit for 500 reactions and choose the VIAFLO 96/384 to add reagents or samples simultaneously.