Fragment analysis for AFLP

If you want to use the multichannel loader, you will have to set up the samples in the correct pattern.

Make up a mixture of size standard and loading dye as follows:

# samples 1 55 110
Tamra size standard (ul) 0.18 10 20
Formamide:dye 5:1 (ul) 1.81 100 200
Total 2 110 220

For each sample, add 2 ul of size/dye mixture to 2 ul of selected PCR. Denature in thermal cycler (90oC 5 min). Put on ice. Load 1 ul per lane on the gel.

Run on 377 using Genescan software. Set up sample sheet first. Then set up Genescan run. Pull in sample sheet. Genescan uses filter set C.

Reagents:

5:1 formamide : loading dye

500 ul formamide
100 ul loading dye (comes with Tamra size standard)

Make up aliquots and keep in freezer.
Tamra size standard

ABI Genescan - 500 p/n 401733