Cemia Services

Overview

Microsatellites, SSRs, and other fluorescently-labeled PCR products can be sized on our ABI 3730xl and ABI 3730 DNA Analyzers. Up to four fluorescent dyes, in addition to the size standard, can be detected. Currently, we run the Applied Biosystems G5 dye set (6-FAM, VIC, NED, PET, and LIZ).

Features

  • 3-5 working days turnaround from sample receipt
  • High quality reads on the ABI3730xl
  • Runs for both tubes and plates

General Considerations

Fragment analysis using Life Technologies products involves:

  • Labeling fragments with fluorescent dyes. Multiple different colored fluorescent dyes can be detected in one sample. One of the dye colors is used for a labelled size standard present in each sample. The size standard is used to extrapolate the base-pair sizes of the sample product peaks.
  • Amplifying the labeled fragments using polymerase chain reaction (PCR) on a thermal cycler.
  • Separating the fragments by size using capillary electrophoresis.
  • Analyzing the data using software to determine Size. The analysis software uses the size standard in each sample to create a standard curve for each sample. It then determines the relative size of each dye-labeled fragment in the sample by comparing fragments with the standard curve for that specific sample.

Submission

Complete all details in the DNA Sequencing Request Form (concentration, type, purification requests etc) and send it via e-mail or enclose it in the parcel. Lack of any information will compromise quality of sequence readout.

On the submission form, provide all necessary contact and charging details.
Use 1.5 ml safe-lock tubes or 96well plates for your samples.
Do not tape or wrap tubes with parafilm. Safe-lock tubes offer perfect sealing and evaporation protection.
Use a water resistant marker for labeling of template and primer tubes.
Check that labeling is clear and simple.
Provide adequate protection within padded envelope.
Do not tape tubes to paperwork or leave loose inside the envelope.


Results

Results can be sent via e-mail.

Electropherograms can be viewed using Editview (Mac platform) or SeqSanner (PC platform) both of which are freely available.