Jan 31, 2015
Version 1
Suggested protocol for loading a DNA Ladder/marker V.1
- New England Biolabs (NEB)Tech. support phone: +1(800)632-7799 email: info@neb.com
External link: https://www.neb.com/protocols/1/01/01/suggested-protocol-for-loading-a-dna-ladder-marker
Protocol Citation: New England Biolabs: Suggested protocol for loading a DNA Ladder/marker. protocols.io https://dx.doi.org/10.17504/protocols.io.cq4vyv
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
Created: January 31, 2015
Last Modified: March 27, 2021
Protocol Integer ID: 508
Keywords: agarose gel, how to load a gel, loading a ladder, loading a marker, DNA ladder, DNA marker, loading a ladder, 1 kb, 100 bp, 1 kb plus
Abstract
This is the suggested protocol for use with λ DNA-Mono Cut Mix (N3019), фX174 DNA-HaeIII Digest (N3026),pBR322 DNA-BstNI Digest (N3031), pBR322 DNA-MspI Digest (N3032), 2-Log DNA Ladder (0.1-10.0 kb) (N3200), 100 bp DNA Ladder (N3231), 1 kb DNA Ladder (N3232), Low Molecular Weight DNA Ladder (N3233), and 50 bp DNA Ladder (N3236)
Before start
The following protocol is recommended for a 5 mm wide lane.
Prepare loading mixture (6 μl total volume):
Note
The components of the mixture should be scaled up or down, depending on the width of the agarose gel.
Distilled water, 4 μl
4 µL
6X Purple no-SDS Loading Dye, 1 μl
1 µL
Gel Loading Dye, Purple (6X), no SDS - 4.0 mlNew England BiolabsCatalog #B7025S
DNA Ladder, 1 μl
1 µL
Mix gently
Load onto the agarose gel
Note
For long term storage, store at -20°C. If samples need to be diluted, use TE or other buffer of minimal ionic strength. DNA may denature if diluted in dH20.