A Technique for Magnetic Glass Particle-Mediated Nucleic Acid Extraction from Blood
Published: November 30, 2023
Abstract
Source: Rosenstierne, M. W., et al. Rapid, Safe, and Simple Manual Bedside Nucleic Acid Extraction for the Detection of Virus in Whole Blood Samples. J. Vis. Exp. (2018).
This video demonstrates a technique for nucleic acid extraction from whole human blood using magnetic glass particles (MGPs) — small magnetic beads coated with silica. Upon lysing the blood cells using a lysis buffer, the released nucleic acids bind to the surface of silica-coated MGPs, extracting the molecules from the sample.
Protocol
All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines 1. Blood Collection from Patients CAUTION: Take appropriate laboratory safety measures when collecting whole blood from the patient. Wear gloves and glasses. If the patient is in isolation, please follow biosafety level 4 procedures. To collect intravenous whole blood from the patient, use a butterfly need…
Açıklamalar
The authors have nothing to disclose.
Materials
| Vacutainer K2 EDTA tubes (4 mL) | Becton Dickinson | 368861 | |
| Plus Blood Collection | Becton Dickinson | 362725 | |
| 23G x 1 needle | Becton Dickinson | 300800 | |
| LUER LOK syringe (3 mL) | Becton Dickinson | 309658 | |
| Butterfly needle with small-bore extension tubing | Jørgen Kruuse A/S | 121714 | |
| 1% Virkon | Nomeco A/S | 849265 | |
| MagNA Pure LC DNA Isolation Kit I – Lysis/Binding Buffer – Refill | Roche Diagnostics A/S | 3246752001 | |
| MagNA Pure LC Total Nucleic Acid Isolation Kit | Roche Diagnostics A/S | 3038505001 | |
| Nunc™ Biobanking and Cell Culture Cryogenic Tubes (1,8 mL) | Thermo Fisher Scientific | 375418 | |
| Nunc™ Biobanking and Cell Culture Cryogenic Tubes (3,6 mL) | Thermo Fisher Scientific | 379189 | |
| Nunc™ Biobanking and Cell Culture Cryogenic Tubes (4,5 mL) | Thermo Fisher Scientific | 379146 | |
| DynaMag™-5 Magnet | Thermo Fisher Scientific | 12303D | |
| Transfer pipette (3.5 mL) | Sarstedt | 86.1171.010 | |
| Thermo Scientific™ Samco™ Fine Tip Transfer Pipettes (1.5 mL) | Thermo Fisher Scientific | 231 | |
| Thermo Scientific™ Nunc™ 50 mL Conical Sterile Polypropylene Centrifuge Tubes | Thermo Fisher Scientific | 339652 |
Bu Makaleden Alıntı Yapın
A Technique for Magnetic Glass Particle-Mediated Nucleic Acid Extraction from Blood. J. Vis. Exp. (Pending Publication), e21842, doi: (2023).