BEFORE YOU BEGIN:
	• Add 4 volumes of ethanol (≥ 95%) per volume of DNA Wash Buffer.
	• All centrifugation steps should be carried out at 16,000 x g (~13,000 RPM).
	• If working with DNA fragments ≥ 10 kb, preheat the appropriate amount of DNA Elution Buffer to 50°C.
	• Please note that the column reservoir holds 800 μl.
PROTOCOL STEPS:
1.	 Excise the DNA fragment from the agarose gel, taking care to trim excess agarose. Transfer to a 1.5 ml microfuge tube,
and weigh the gel slice. Minimize exposure to UV light.
2.	 Add 4 volumes of Gel Dissolving Buffer to the gel slice (e.g., 400 μl buffer per 100 mg agarose). If the gel slice is > 150 mg,
consider reducing the amount of Gel Dissolving Buffer to 3 or 3.5 volumes to minimize the guanidine salt present in the workflow.
3.	 Incubate the sample between 37-55°C (typically 50°C), inverting periodically until the gel slice is completely dissolved
(generally 5-10 minutes). For DNA fragments > 8 kb, an additional 1.5 volumes of water should be added after the slice is
dissolved to mitigate the tighter binding of larger pieces of DNA (e.g., 100 μl gel slice: 400 μl Gel Dissolving Buffer: 150 μl water).
4.	 Insert column into collection tube and load sample onto the column. Spin for 1 minute, then discard flow-through.
5.	 Re-insert column into collection tube. Add 200 μl DNA Wash Buffer and spin for 1 minute.
Discarding flow-through is optional.
6.	 Repeat step 5.
NEB #T1020
continued on back →
For a detailed protocol, or to download the full manual, visit www.neb.com/ T1020.
Monarch
®
DNA Gel Extraction Kit Protocol Card
7.	 Transfer column to a clean 1.5 ml microfuge tube. Use care to ensure that the tip of the column
does not come into contact with the flow-through. If in doubt, re-spin for 1 minute.
8.	 Add ≥ 6 μl of DNA Elution Buffer to the center of the matrix. Wait for 1 minute, and spin for
1 minute to elute DNA. Typical elution volumes are 6-20 μl. Nuclease-free water (pH 7-8.5) can also
be used to elute the DNA. Yield may slightly increase if a larger volume of DNA Elution Buffer is used, but
the DNA will be less concentrated. For larger size DNA (≥ 10 kb), heating the elution buffer to 50°C prior
to use can improve yield.
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc. For more information about commercial rights,
please email us at busdev@neb.com. The use of this product may require you to obtain additional third party intellectual property rights for certain applications.
New England Biolabs is an ISO 9001, ISO 14001 and ISO 13485 certified facility.
© Copyright 2020, New England Biolabs, Inc.; all rights reserved.
This card is made with FSC certified 100% post-consumer fiber. Please recycle.
Want to use this kit to purify DNA from PCR
and other enzymatic reactions?
Simply purchase the Monarch DNA Cleanup Binding Buffer
(NEB #T1031L) and use with this kit. Protocol available
at www.neb.com/T1030
Questions?
Our tech support scientists would be happy to help.
Email us at info@neb.com
V5.0 – 1.21 #101028