Luckau Protocols:1Kb Ladder

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1Kb Ladder Mix

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Contents

Purpose

Nucleic acid ladder is used as a size standard in agarose gel electrophoresis. It contains DNA molecules of known lengths. The ladder is pre-mixed with gel load buffer (color dyes with varying mobility on an agarose gel) for direct addition of PCR product (or genomic DNA) and transfer to the gel.


Materials

Invitrogen 1Kb Ladder

Promega Blue/Orange Loading Dye, 6x

  • catalog# G1881
  • use at 1x concentration

Other Knowns

  • well width (24-tooth comb) = 6mm


Protocol

Calculations

  • 6mm x (0.1µg ladder)/(1mm width) x (1µL/1µg) = 0.6µL ladder per well
  • Per well: 0.6µL ladder + 4.4µL H2O + 1µL loading dye = 6µL total ladder mix

Make Ladder Mix

120µL ladder + 880µL H2O + 200µL loading dye = 1200µL ladder mix

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