Preparation of DNA

P rotocol
Ladders/Markers for Electrophoresis

This protocol is for the Preparation of DNA Ladder/Markers for Electrophoresis

Table 1. Recommendations for loading the conventional formulation (supplied in TE buffer) DNA ladders/markers.

Conventional formulation (supplied in TE buffer) DNA ladders/markers

Technical specifications
Lambda Phage & Plasmid Markers for Genomic
GeneRuler™ DNA ladders
DNA markers DNA markers DNA Analysis
50 µg (100 µl) is sufficient for: 50 µg (100 µl) is 50 µg (100 µl) is sufficient for: 6 µg (30 µl) is
Supplied amount / sufficient
• 100 applic. on agarose gel • 100 applic. on agarose gel sufficient for 120 applic.
number of applic. for 100 applic.
• 50 applic. on native PAGE • 50 applic. on native PAGE on agarose gel
on agarose gel
Amount used per 0.1 µg (0.2 µl) for agarose gel 0.1 µg (0.2 µl) 0.1 µg (0.2 µl) for agarose gel
6 ng
1 mm width of a gel lane 0.2 µg (0.4 µl) for PAGE for agarose gel 0.2 µg (0.4 µl) for PAGE
Mix 1 µl (0.2 µg) of
Dilution Not needed Not needed Not needed DNA marker with 39 µl
of nuclease-free water
Heat at 65°C Heat at 65°C for 5 min;
for 5 min;
Heating Do not heat Do not heat chill on ice for 3 min
chill on ice for 3 min
before use before use
I. Loading on agarose gel:
DNA ladder/marker loading 10 µl (50 ng) of
1 µl (0.5 µg) 2 µl 9 µl 1 µl (0.5 µg) 2 µl 9 µl 1 µl (0.5 µg) 2 µl 9 µl
dye Water, nuclease-free diluted marker 1 µl
Mix gently and load on gel
II. Loading on polyacrylamide gel:
DNA ladder/marker loading Not recommended Not recommended
2 µl (1 µg) 0.5 µl 0.5 µl 2 µl (1 µg) 0.5 µl 0.5 µl
dye Water, nuclease-free for PAGE for PAGE
Mix gently and load on gel
II. Loading on polyacrylamide gel:
DNA ladder/marker loading Not recommended Not recommended
2 µl (1 µg) 0.5 µl0.5 µl 2 µl (1 µg) 0.5 µl 0.5 µl
dye Water, nuclease-free for PAGE for PAGE
Mix gently and load on gel
Table 2. Recommendations for loading ready-to-use DNA ladders/markers.

DNA ladders/markers, ready-to-use

Technical
GeneRuler™ & ZipRuler™ Phage &

P rotocol
specifications MassRuler™ FastRuler™ O’RangeRuler™ Lambda DNA
O’GeneRuler™ Express DNA Plasmid DNA
DNA ladders DNA ladders DNA ladders markers
DNA ladders ladders markers

Supplied 2x500 µl for 2x500 µl

500 µl for 100 2x500 µl for 500 µl for 500 µl for 500 µl for
volume/number 50-333 for 100-200
applic. 50-200 applic. 100 applic. 100 applic. 100 applic.
of applic. applic. applic.
Heat at 65°C for
5 min;
Heating Do not heat Do not heat
chill on ice for
3 min before use
Mix gently and load on gel
Volume per 1
mm width of a 1-2 µl variable variable 1-2 µl 1-2 µl 1-2 µl 1-2 µl
gel lane

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