Microsatellites/IDAA/FLA/STR
Fragment Length Analysis Tailored To Your Needs

You run the PCR, we do the rest!

Convenient fragment length analysis (FLA) service

Just submit your PCR products. Experts take care of the subsequent fragment length analysis steps that are included into the standard FLA services.

The fragment length analysis includes the following services:

• Test run with a small number of samples
• Normalisation of PCR product concentration* and addition of loading dye and size standard
• Accurate separation of the fragments by capillary electrophoresis on ABI 3130 XL sequencing machine
• Comprehensive data analysis
• Results sent by email as XLS and FSA files

Matrix, filter sets and dye combinations are described below.

* For multiplex analysis, the concentrations of the PCR products in a sample must be normalized before shipment in order to get a balanced profile (variations of max. 50% are recommended).

You can order this product online, using the "order now" button on top!

Your fragments are fully prepared?

Make use of the ready-to-load service!

Ready-to-load fragment length analysis (FLA) service

The ready-to-load FLA includes the following services:

• Accurate separation of fragments (100-550 bp) by capillary electrophoresis (CE) on ABI 3130 XL sequencing machine under standard conditions
• Results send by email as FSA files (GeneMapper or Genotyper analysis software required)
• Larger fragments on request
  
  

Matrix, filter sets and dye combinations are described below.

You can order this product online, using the "order now" button on top! 

Tailor your individual FLA project. Benefit from expertise in DNA extraction, marker selection and assay design.

Send us your request. (see button on top)

Use our fragment length analysis service for your Indel profiling in CRISPR experiments as described by Yang et. al (2015).

You can order this product via our Excel order form (see button on top).

Sample Data and Formats

• You can send your samples in following formats:
  • 0.2 ml tubes (up to 8 samples per batch)
  • 8-well strips (up to 32 samples per batch)
  • Non-flexible 96-well PCR plates, preferably semi-skirted with V-well bottoms
• Please use 96-well plates whenever possible but especially for sample numbers of at least 33 samples per batch.
• Please use the following 96-well plate type especially for Ready-to-Load samples: e.g. MicroAmp® Optical 96-Well Reaction Plate (Thermo Fisher Scientific Inc., cat. no. 4316813)

Sample preparation and concentration

• Label your samples according to the sample names indicated in your sample list:
  • Single tubes on the lid o 8-strips on the side
  • 96-well plates on the plate frame
• Concentration across all samples must be normalised
• For multiplex analysis, the concentrations of the PCR products in a sample must be normalised, in order to get a balanced profile (variations of max. 50% are recommended)
• The concentration of each PCR product resp. marker should be 50 – 100 ng/µL
• Use 8-cap strips for sealing 8-well strips and 96-well plates (recommended)
• Alternatively, adhesive PCR foil (e.g. from 4titude® , code: 4ti-0500) can be used
  • Make sure the plate has reached room temperature and the plate surface is absolutely dry before you stick the foil on the plate
  • Seal the plate tightly and avoid folding; don’t freeze and thaw the plate after sealing

FLA Ready-to-Load Service

• Perform your PCR with the respective dye labelled primers which can be analysed with one of the above described filter sets
• The concentration of the PCR products should be 50 – 100 ng/µL
• Mix the Ready to Load samples as follow:
  • 10 µL Hi-Di Formamide loading buffer (Applied Biosystems)
  • 0.5 µL internal size standard (e.g. ROX-500 or LIZ-500).
  • 2 µL per PCR product
• The total sample volume should be 10 – 15 µL.
• Delivery Time: usually, 5 days.

Fragment Length Analysis Service

• Perform your PCR with the respective dye labelled primers which can be analysed with one of the above described filter sets
• Send us the PCR products in a concentration of 50 – 100 ng/µL
• The total sample volume should be 10 – 20 µL.
• Delivery Time: usually, 10 days

Matrix, Filter Sets and Dye Combinations

Following filter sets and dye combinations are used as matrix and for labelling the PCR products:

• FAM, NED, HEX and ROX* (ABI Standard Dye Set DS-30)
• FAM, JOE or HEX, TAMRA and ILS600 (Promega ILS600)
• FAM, VIC, NED and ROX* (ABI Standard Dye Set DS-31)
• FAM, JOE, NED and ROX* (ABI Standard Dye Set DS-32)
• FAM, VIC, NED, PET and LIZ* (ABI Standard Dye Set DS-33)
• FAM, Yakima Yellow, ATTO 550, ATTO 565 and ATTO 663* (Eurofins alternative dye set EF-01)

*the dyes ROX, LIZ are reserved for the size standard and should not be used for PCR product labelling.

Additionally, also dye ATTO 663 should not be used as it is detected in the same channel like LIZ size standards.

Possible size standards that can be added from our side are: ROX-500, LIZ-500, ILS600 as well as LIZ1200. The Eurofins alternative dye set EF-01 can only be combined with LIZ size standards.

You can ship your samples at room temperature.

You have 2 shipment options:

• For convenient sample shipment you can use our dropboxes. Please note that the delivery will take 1 day more, so if you are in a hurry we recommend direct sample shipment'
  
  
• Direct sample shipment

Eurofins Genomics FEB

Anzingerstrasse 7a

85560 Ebersberg

Germany