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96-WELL SEQUENCING SERVICE 

For routine sequencing analysis of large numbers of samples, BaseClear offers a special high throughput service. Our state-of-the-art robotics and automated protocols guarantee fast and accurate results, and offers a significant cost reduction compared to single run sequencing.

BaseClear's 96-well sequencing service: 

  • Samples can be delivered in either half (up to 47 samples) or full (up to 95 samples) plates 
  • You can use any 96-well plate (e.g. standard PCR plate) 
  • Please leave well H12 empty, this well will be used for internal sequence control
  • Single sequencing reaction with a run-time corresponding to either 1000 bp (long run) or 550 bp (short run) 
  • Sequencing data are automatically analysed by KB caller, the international standard software for superior base calling and sequence data interpretation 
  • Sequence results are sent directly to the customer in FASTA, SCF and ABI format
  • Re-runs are possible upon request
  • Delivery time: 1-2 working days
  • Make sure that the plate is properly sealed and packaged to prevent leakage or damage during transport

SAMPLE DELIVERY WHEN SAMPLE PLATE AND PRIMERS ARE SENT IN SEPARATE

  • 96-well plate with bacteria either as glycerolstocks on dry ice or as cultures in LB medium. 
  • 96-well plate with purified plasmid in a minimal volume of 20 ul per well with a DNA concentration of 50 – 200 ng/ul. Make sure the DNA concentrations are normalised over the plate.
  • 96-well plate with purified or Raw PCR product in a minimal volume of 20 ul per well with a DNA concentration >5 ng/μl. Make sure the DNA concentrations are normalised over the plate. 2 μl of the PCR product on agarose gel should give a single clear and sharp band.

DNA preferably column purified dissolved in water or 10mM Tris pH8.0. Please note that DNA samples should be free of EDTA and ethanol since trace amounts of these compounds will inhibit the sequence reaction.
 

Primers

You can choose a maximum of one primer per half plate which will be added by BaseClear. If you want to use multiple primers per plate, you can submit a ‘mirror-plate’ with the corresponding primers.

  • For a full 96-well plate please sent in 200 ul primer with a concentration of 10 pmol/ul.
  • For a half 96-well plate please sent in 100 ul primer with a concentration of 10 pmol/ul
  • For a mirror-plate please add 20 ul primer with a concentration of 10 pmol/ul per well in a 96-well plate.
 

96-WELL PREMIX: PURIFIED DNA PREMIXED WITH PRIMER

  • For purified plasmid samples, submit 0,75 µg plasmid DNA mixed with 25 pmol of primer in a total volume of 20 µl per well
  • For PCR product samples, submit 15 ng per 100 bp PCR product with 25 pmol of primer in a total volume of 20 µl per well

Meet Leon de Boer!

Leon is manager of our Sanger sequencing department and responsible for the quality assurance and the delivery of data at the agreed upon times. He always runs a final check before data are sent off to the customers.

Testimonial Erasmus MC Rotterdam

BaseClear sequenced for us thousands of samples of people suffering from severe mental and physical disabilities. I have used that data for my PhD thesis research to investigate if there was a relation with mutations in certain thyroid hormone transporters or enzymes. With BaseClear sequencing all the DNA, I was able to devote my time to all the other aspects of the research. The guidance and support was very positive not to mention the technical assistance that really addressed the needs we had.

PostDoc Internal Medicine

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