Oct 11, 2021
Version 2
DNA Ethanol Precipitation (SOP009.v1.1) V.2
This protocol is a draft, published without a DOI.
- 1Arizona State University
- Human Cell Atlas Method Development Community
Protocol Citation: Rory Kruithoff, Douglas Shepherd 2021. DNA Ethanol Precipitation (SOP009.v1.1). protocols.io https://protocols.io/view/dna-ethanol-precipitation-sop009-v1-1-byxupxnwVersion created by Rory Kruithoff
Manuscript citation:
From Qiagen: https://www.qiagen.com/it/resources/faq?id=5d591b8b-968a-4a17-849f-9d0f719b40af&lang=en&Print=1
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: Working
We use this protocol and it’s working
Created: October 11, 2021
Last Modified: October 11, 2021
Protocol Integer ID: 53972
Keywords: DNA, ethanol, ethyl alcohol, precipitation,
Abstract
Document Summary: This document, SOP002 - DNA Ethanol Precipitation, describes a method to concentrate or dry DNA using ethanol to reduce the solubility of dissolved DNA causing it to precipitate out of solution. Alternatively, a vacuum concentrator can be used if one is available.
Attachments
Materials
Required Reagents
- Sodium AcetateSigmaCatalog #S2889-250g
- Ethanol 99.5% ACS ReagentThermofisherCatalog #AC615090010
- Nuclease-free water
Required Equipment
- Microcentrifuge
- Sterile Eppendorf tubes
- -20°C Freezer
Safety warnings
For hazard information and safety warnings, please refer to the SDS (Safety Data Sheet).
Before start
Quick Overview:
Step 1 – Precipitate DNA
Step 2 – Pellet DNA and dry
Step 3 – Resuspend DNA
v1.1 revision notes
- Updated SOP to standard formatting
- Updated document description
Precipitate DNA
Precipitate DNA
2h
2h
Add 1/10 volume of 3 M Na-Acetate 5.2 , and 2 to 2.5 volumes of ice-cold 100% ethanol to the DNA sample.
Mix, and store at -20 °C for at least 01:00:00 to precipitate the DNA. Typically, DNA will be left at -20 °C Overnight before proceeding to part 2.
2h
Pellet DNA and dry
Pellet DNA and dry
35m
35m
Recover the precipitated DNA by centrifugation at full speed in a microcentrifuge for 00:15:00 - 00:20:00 to form a pellet.
20m
Pour off the ethanol and wash the pellet twice with Room temperature 70 % (v/v) ethanol .
Allow the DNA pellet to air-dry.
Resuspend DNA
Resuspend DNA
Re-suspend the DNA in a suitable volume of sterile TE buffer or nuclease-free water.
Store at -20 °C and avoid any unnecessary freeze-thaw cycles.