Fragment Analysis

Fragment analysis with GeneScan ROX size standard

First time users

Please contact the LGTC at info@lgtc.nl before submitting your first plate.  The peak height of your results depends on the concentration of your PCR product. The concentration of your PCR depends on the amplification percentage of your marker/primer. Ideally, you should obtain peak heights of 2000-4000. Due to large differences in the amounts of markers being used it is not possible to recommend one specific dilution for the ABI3730xl runs. Previous tests have shown that some amplified material needed to be diluted up to 200-fold, depending on the marker that was used. To help you to obtain the best results, we advise you to start with a test plate (96 samples) with scaled up dilutions of known samples, amplified with different markers of your list. You should start with dilutions from 50x up to 200x of the amplified material and then work down.   

Sample delivery

Samples should be ready-to-run for analyzing by the ABI3730xl. Deliver them in a 96-well skirted plate (preferably MicroAmp Optical 96-well Reaction Plate Applied Biosystems Part# 4306737). We have barcode stickers if needed.  Please contact the LGTC at info@lgtc.nl when you have samples ready to run.

Fragment analysis services

  • 4 dye Fragment Analysis with ROX as size standard.
  • 384-well plates ROX as size standard.

We accept full 96 well plates but if you do not have 96 samples be sure to fill the empty wells with MQ.

Data

Fragment run results (fsa files) should be available for collection within 3 working days. You will receive a mail with a zip file containing the results.