Luckau Protocols:1Kb Ladder

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.

Invitrogen 1Kb Ladder

 * catalog# 15615-016
 * Invitrogen says to use: 0.1µg ladder / mm width of well
 * STK at 1.0µg/µL
 * [[Media:Invitrogen_1KbLadderProductManual.pdf]]

Promega Blue/Orange Loading Dye, 6x

 * catalog# G1881
 * use at 1x concentration

Other Knowns

 * well width (24-tooth comb) = 6mm

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