Individual Tube Submission Policies

  1. All samples must be submitted with primer and template premixed, except for primer walk projects.
  2. We only accept individual tubes submitted in VWR Labcon and American Bioanalytical purple screw cap tubes. The American Bioanalytical tubes are located in both Yale stockrooms. Individual tube premixed reaction submitted in any other tube types will not be accepted. The American Bioanalytical tubes can be order by phone at 1-800-443-0600 or through email at The catalog number is PL73310-00001 and they are packaged 500 tubes with caps.
  3. Due to our sequencing methods all individual tube samples should be labeled with barcodes prior to submission. Barcodes are given out over the counter in the Kline and Medical Stockrooms at no additional cost. Please see Barcode Labeling Instructions for more information.

Individual Tube Submission Protocol

Provide primer and template premixed according to the guidelines below. We offer a high volume service for large projects of 48 or more samples submitted at a time. Please see High Volume Submission Policies and Procedures [ST1] for more information.

The protocols below are sufficient for two sequencing reactions (e.g. 300ng DS DNA/sequencing reaction) with one template/primer combination. This allows us to repeat a sample in case of technical failure without having to contact you for more sample.

DS plasmid DNA:

  • 500-600 ng ds plasmid DNA template
  • 2 µl 4 µM primer
  • x µl sterile water
  • 18 µl total volume

PCR Product:

  • 10-20 ng template for every 200 bases of PCR fragment length
  • 2 µl 4 µM primer
  • x µl sterile water
  • 18 µl total volume

e.g. 500bp PCR fragment, mix 25-50 ng PCR product with 2 µl 4 µM primer and sufficient water to bring the final volume to 18 µl.

Cosmid, BAC and Phage Lambda DNA:

  • Mix as above except add 2000-4000 ng template DNA.
  • Specify Cosmid, BAC or Phage DNA on sample sheet so we will use altered cycling condition.
  • Only high purity templates should be used to optimize the amount of DNA loaded onto the sequencer.
  • BAC template preparation methods that we recommend:
    1. Alkaline lysis with phenol extraction and isopropanol precipitation.
    2. Cesium Chloride Banding

Single-stranded M13 or Phagemid DNA:

  • Mix as above except add 200-300 ng template DNA.

Standard Turn Around Time

Our turnaround time is 24 - 36 hours from the time we receive your samples (except for samples received on Friday). For example, samples received in the morning by 10:30 am will be returned the next business day before noon.