GMT Tutorial Exercise 21 - Workflow 1 - Step 5

Using the macro - the Primer Dilution Plate Layout worksheet

• Users of the Whole Genome Scan kits are provided with ready-to-use primer aliquots, located on a number of 96-well plates.
• The Primer Dilution Plate Layout worksheet of the macro gives details where primer mixes are found on these 96-well plates, and how they have been made up. Use this information if e.g. having to make up some of the primer stocks. For every well of several "Dilution Plates", the following information is shown (see the highlighted example in the screenshot below):
  • Name of the "locus-specific" primer pair in a well (e.g. ta0185)
  • Volume (in µL) of 0.4 µM primer mix to be pipetted into the well, if the diluted primer stock needs to be made up (first number, e.g. 210 µL). (The "0.4 µM primer mix" is a mix of the forward and reverse locus-specific primers, with both primers at a concentration of 0.4 µM.)
  • Volume (in µL) of 10 mM Tris-HCl, pH 8, to be pipetted into the well, if the diluted primer stock needs to be made up (second number, e.g. 390 µL)
• Primers have been prepared in the specified layout to simplify mixing "locus-specific" primers and PCR master mixes for the different DNA samples by manual or robotic multichannel pipetting, while ensuring that "marker panels" are not mixed up.
• Click on Print Preview to print a copy for use during PCR set-up.
• Click on Next Sheet to begin the [next step] of the workflow.