PCR fragment analysis:
Any amplified labeled at 5 'that has a length between 50bp and 560bp can be analyzed with the sequencer.
The advantages are:
- higher resolution and sensitivity than agarose gel
- possibility of having the precise measurement of the length of the fragment in reference to an internal molecular weight standard (liz 600) which is run together with the samples.
- possibility to have a relative quantitative measure based on the amplitude of the peak and the intensity of the signal.
- possibility to analyze many samples at the same time.
How to prepare the samples to send:
- 10ul of amplification for each marker (indicate on the form, in the additional information, the expected measurements, base pair, for each sample)
- ship at room temperature in 0.2ml tubes
- it is recommended to verify the amplifiers and the quantifications on agarose gel before sending the samples
PCR amplifiers deriving from different protocols and techniques (Microsatellites, AFLP, OLA, SNPLex, SNAPShot etc.) must be obtained using one or more of the following markers: -6-FAM, VIC, NED and PET.
The results are sent in 24/48 hours and summarized in an Excel sheet with the measurement of the fragments, the area of the peaks and the data points with the corresponding plot images.
Genotyping of laboratory animals:
The service allows the identification and characterization of laboratory animals and includes:
- the extraction of genomic DNA from biopsies
- synthesis of oligos to amplify gene (s)
- agarose gel (on request the sequencer can be used to analyze the amplification profiles)
How to prepare the samples to be sent:
- in Ethanol: use single 1.5 ml tubes identified with progressive number
- in dry ice: use single 1.5 ml tubes identified with progressive number or 96-well deep-well with relative identification scheme
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