Sample Submission Guidelines for Fragment Analysis

Eurofins Genomics (US) offers the ‘Fragment Analysis – Ready2Load’ service using ABI 3730xl DNA analyzers. Our workflow is optimized to offer the service for the PCR products generated by microsatellite, SNP, LOH, AFLP, RFLP, T-RFLP, ARISA or other protocols.

Dye Set

Only the following dye combinations can be used as matrix and for labelling the PCR products:
ABI Standard Dye Set DS-336-FAM, VIC, NED, PET and LIZ*
*the dye LIZ is typically reserved for the size standard and should not be used for PCR product labelling.Size Standard

The protocol for the Ready2Load fragment analysis at Eurofins Genomics US is optimized for analysis of DNA fragments 50–600 bp. The LIZ 600 size standard must be included in your samples for fragment analysis.

Sample preparation for ‘Fragment Analysis – Ready2Load’ Service

Please adhere to the following guidelines, to assemble samples for Fragment Analysis – Ready2Load service.

• Run PCR using dye labeled primers that can be analyzed with ABI Standard DS-33 filter set
• Prepare solutions of PCR products at 50–100 ng/μL
• Assemble the samples for ‘Fragment Analysis – Ready2Load’ Service, by mixing the following:
  1. Hi-Di Formamide loading buffer, 10 μL (Applied Biosystems)
  2. Dye-Labeled internal size standard, 0.5 μL (use only LIZ-600).
  3. PCR product, 2 μL
• The total sample volume should be 10–15 μL.

Sample Submission formats

Please note that you can submit your samples in one of the following two formats:

• 1.75 mL centrifuge tubes
• 96-well plates (MicroAmp® Optical 96-Well Reaction Plate, ThermoFisher, catalogue no. 4316813, or equivalent)

Shipping

Ship the liquid samples at ambient temperature to:

ATTN: Fragment Analysis Sequencing Lab
Eurofins Genomics
12701 Plantside Drive,
Louisville, KY 40299

• Use 8-cap strips for sealing 96-well plates (recommended)
• Seal the plate tightly and avoid folding; don’t freeze and thaw the samples after sealing.