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DNA Sequencing and Synthesis Facility at CEMB
 
       

DNA Sequencing using the ABI PRISM - 3100

Genetic Analyzer

 

The ABI PRISM 3730 & 3100 Genetic Analyzers are state of the art machines for automated DNA capillary electrophoresis sequencing & genotyping. Any primer can be used (provided by the customer), and we will guarantee more than 800 bases of good quality fluorescence-based DNA sequence. As with all sequencing, the quality of the sequence is dependent on the quality of the template DNA and primer as well as careful quantitation of the DNA concentration.

Samples will be accepted in the form of highly purified DNA template

 

Templates should be provided at the following concentrations:

 

Template

Concentration

Plasmid

300 ng/ml

Cosmid

1 mg/ml

BAC

1 mg/ml

Bacterial Genomic

3 mg/ml

Phage

1 mg/ml

 

Template

Concentration

PCR Fragment   100 - 200 bp

3 ng/ml

200 - 500 bp

10 ng/ml

500 - 1000 bp

20 ng/ml

1000 - 2000 bp

40 ng/ml

> 2000 bp

100 ng/ml

 

Primers should be provided at a concentration of 3.2 Pico mol/ul 

Total quantity required

Template                       5.0ul

Primer                           5.0ul

 

  • Estimated time for results will be 4-5 working days and results will be given via e-mail or in the form of print outs of electropherograms. Soft copies of data on customer’s CD can also be provided if requested.

 

The electopherograms can be visualized using the following programs:

 

  • ABI Sequencing Analysis (Windows)
  • BioEdit  (Windows)

·        Chromas 1.45 (Windows)

Manual editing of the sequence:
We manually verify computer-generated base calling to ensure the highest sequencing accuracy.


 
 

DNA Core Facilities

 
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