Suggested protocol for loading a DNA Ladder/marker
New England Biolabs
Abstract
This is the suggested protocol for use with:
Quick-Load®Purple 1 kb DNA Ladder (N0552)
Quick-Load®Purple 100 bp DNA Ladder (N0551)
Quick-Load®Purple 50 bp DNA Ladder (N0556)
Quick-Load®1 kb Extend DNA Ladder (N3239)
Quick-Load®1 kb DNA Ladder (N0468)
Supercoiled DNA Ladder (N0472S)
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)
50 bp DNA Ladder (N3236)
Before start
This protocol is recommended for a 5 mm wide gel lane. The components of the mixture should be scaled up or down, depending on the width of the lane.
Steps
Prepare loading mixture ( 6 μl total volume):
| A | B |
|---|---|
| Distilled water (dH20)* or TE Buffer | 4 μl |
| Gel Loading Dye, Purple (6X), no SDS | 1 μl |
| DNA Ladder/Marker | 1 μl |
| Total Volume | 6 μl |
*For multiple loads, dilution, and storage, use TE or other buffer of minimal ionic strength instead of water. DNA may denature if diluted and stored in dH20.
Mix gently by pipetting.
Load onto the agarose gel.
