Jul 29, 2024
Version 1
Opentrons Pipeline: DNA Extraction with the Mag-Bind Blood & Tissue DNA HDQ 96 Kit Tissue Protocol V.1
- Thalía Silvestre1
- 1UNMSM
Protocol Citation: Thalía Silvestre 2024. Opentrons Pipeline: DNA Extraction with the Mag-Bind Blood & Tissue DNA HDQ 96 Kit Tissue Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l62rqrgqe/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: In development
We are still developing and optimizing this protocol
Created: July 27, 2024
Last Modified: July 29, 2024
Protocol Integer ID: 104196
Keywords: Opentrons, OT-2, Mag-Bind Blood & Tissue DNA HDQ 96 Kit Tissue Protocol
Disclaimer
DISCLAIMER – FOR INFORMATIONAL PURPOSES ONLY; USE AT YOUR OWN RISK
The protocol content here is for informational purposes only and does not constitute legal, medical, clinical, or safety advice, or otherwise; content added to protocols.io is not peer reviewed and may not have undergone a formal approval of any kind. Information presented in this protocol should not substitute for independent professional judgment, advice, diagnosis, or treatment.
Abstract
This protocol is an automated pipeline to extract a full 96-well plate of DNA from tissue lysates.
This protocol was developed and optimized for the following:
Platform: Opentrons OT-2 automated pipetting robot
Kit: Mag-Bind‱ Blood & Tissue DNA HDQ Prefilled 96 Kit
Recommended number of samples: 96
Two ellutions: Firts ellution( 60μL ) , Second ellution ( 30μL)
Samples Lysis
Samples Lysis
Preparate 96 samples lysis following the Mag-Bind‱ Blood & Tissue DNA HDQ 96 Kit Tissue Protocol:
Mince up to 10 mg tissue and transfer to a 1.5 mL tubes (one for each sample).
(Cut each 10 mg sample with sterilized cutters as small as possible)
Add 250 μL TL Buffer to each sample.
Add 20 μL Proteinase K Solution to each sample. Vortex to mix.
Incubate at 55°C overnight in a shaking water bath.
Centrifuge at maximum speed (≥4,000 x g) for 5 minutes to pellet undigested tissue
debris and transfer 200 µL of the supernatant to the VWR 96-Well Deep Well Plate (1000 μL) without
disturbing the undigested pellet
List of materials to start to use OT-2
List of materials to start to use OT-2
It is necessary to have this equipment to use this pipeline.
Opentrons Equipment List:
| A | B | |
| OT-2 | ||
| OT-2 8-Channel Pipette P300 | ||
| OT-2 Magnetic Module GEN2 | Slot 7 |
Tips & Labware:
| A | B | |
| Four Opentrons 200µL Filter Tips | Slot: 5,6,8,9 | |
| Two Nest 1-Well Reservoirs (195 mL) | Slot: 10 and 11 | |
| VWR 96-Well Deep Well Plate (1000 μL) | Slot 7 on the Magnetic module | |
| Two Nest Well plate ( 100 μL) | Slot 2 and 3 | |
| Two Nest 12 Well Reservoirs (15 mL) | Slot 1 and 4 |
Star OT-2 run
Star OT-2 run
The pipeline start on the step number 7 of the Mag-Bind‱ Blood & Tissue DNA HDQ 96 Kit Tissue Protocol
Load the pipeline on the Opentrons app: 
Extracción.py11KB
Extracción.py11KB The VWR 96-Well Deep Well Plate (1000 μL) labware can be download from: dx.doi.org/10.17504/protocols.io.dm6gp39njvzp/v1
Order of the reagents on the Two Nest 12 Well Reservoirs (15 ml)
Slot 1:
| A | B | C | D | E | F | G | H | I | J | K | L | |
| Well 1 | Well 2 | Well 3 | Well 4 | Well 5 | Well 6 | Well 7 | Well 8 | Well 9 | Well 10 | Well 11 | Well 12 | |
| 11.5 mL of AL buffer | 11.5 mL of AL buffer | Mag-Bind mixed with HBQ buffer (20:340) in a total of 12 mL | Mag-Bind mixed with HBQ buffer (20:340) in a total of 12 mL | Mag-Bind mixed with HBQ buffer (20:340) in a total of 12 mL | 15 ml of VHB buffer | 15 ml of VHB buffer | 15 ml of VHB buffer | 15 ml of VHB buffer |
Slot 4:
| A | B | C | D | E | F | G | H | I | J | K | L | |
| Well 1 | Well 2 | Well 3 | Well 4 | Well 5 | Well 6 | Well 7 | Well 8 | Well 9 | Well 10 | Well 11 | Well 12 | |
| 15 ml of VHB buffer | 15 ml of VHB buffer | 15 ml of VHB buffer | 15 ml of VHB buffer | 15 ml of SPM buffer | 15 ml of SPM buffer | 15 ml of SPM buffer | 15 ml of SPM buffer | Ellution Buffer |