Sample requirements

For Fragment analysis, here is some information to help plan your experiment:

• We expect to receive the sample plate ready to be loaded into the instrument
• Dye set compatible with our instrument: DS-30/D (6-FAM/HEX/NED/ROX) and DS-33/G5 (6-FAM/VIC/NED/PET/LIZ). 
• Standard selection depends on the size of your fragment, the standard generally used is the LIZ 500 but other standard are available. For more information about this, see page 41 of this guide.
• Plates recommended for this application are: Applied Biosystems MicroAmp Optical 96-well plates (Ref. N8010560) or StarLab 96-well Semi-skirted with raised rim plates (Ref. E1403-8200)
• Plate can be seal with adhesive
• Processing depends on when the plate is received, it will be either on the day or the following day
• Raw data file (.fsa) are the standard output provided

Each sample should be prepared as follow, we request 20μl for processing:

Prepare the formamide:size standard mix using:
• 1000 μL of Hi-Di™ Formamide (P/N 4311320) or similar quality formamide
• 50 μL of GeneScan™ 500 LIZ 
For loading, mix 0.5 μL of pooled PCR products with 9.5 μL of formamide:size standard mix.
Please use 1μl of pooled PCR product with 19μl of formamide:size sandard mix for a final volume of 20μl. 
We request 20μl in case of evaporation or sample loss during transit/before processing
Note that this assume that you have good yield from your PCR reaction.