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High Volume Submission Policies

High Volume Submission Policies

The high volume, 96-well plate format is available for researchers who are experienced users of the facility and are confident in their methodology for template and primer preparation. We sequence and charge for all samples submitted on the plate regardless of the results, unless it is a clear technical error on our part.

  • All high-volume plate submissions must be in one of the following plates and sealed carefully with strip caps or sealing foil:

o 96-well Eppendorf twin tec plates Part#951020389,951020346, 951020320, 951020362 (Strip caps Part# 951023035)

o Applied Biosystems Optical 96-well reaction plates Part# N801-0560 (Strip Caps Part# N801-0534

o DOT Scientific PCR plates Part # 351-PCR, 351LP-PCR, 352-PCR, 354-PCR and 650-PCR (Strip Caps Part#405-12PCR)

**The plates are located in the Yale stockrooms or they can be ordered on SciQuest**

**Sealing foil can be purchased from DOT Scientific, Part #T392.

The minimum number of samples for a single high volume plate is 48. When submitting less than 96 samples on a plate, please submit in multiples of 12- placing the samples in rows of 12 consecutively i.e. begin at A1 through A12 then B1-B12, C1-12, etc.

  • High volume plates should be dropped off with a high-volume submission confirmation sheet (available to print after web ordering via our “Sample Submission” page).

The protocols below are sufficient for multiple sequencing reactions 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:

  • 1,000-1,500 ng ds plasmid DNA template
  • 2 µl 4 µM primer
  • x µl sterile water
  • 18 µl total volume

PCR Product:
  • Template
    • 100-300bp:40-50ng
    • 300- 500bp:40-160ng
    • 500bp- 1kb:500ng
  • 2 µl 4 µM primer
  • x µl sterile water
  • 18 µl total volume
  • ***Nanodrop: Please be sure your initial readings are under 1. Anything over 1 is not accurate and will hinder the sequencing process. If they are over 1, dilute the sample until it is in an accurate range.***

  • ***PCR Cleanup: Please be sure samples are cleaned up prior to submission, not just diluted or desalted. Size exclusion columns are recommended. EtOH precipitation is not recommended as this does not remove unbound dNTPs and primer.***

BAC and Phage Lambda DNA:
  • Mix as above except add 4000 ng template DNA.
  • Specify BAC or Lambda when ordering.
  • Only high purity templates should be used to optimize the amount of DNA loaded onto the sequencer.
  • Recommended BAC template preparation methods:
  • 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.
  • Cap the plate firmly using the strip caps or sealing film and wrap each plate with parafilm. (Sealing film is preferable to strip caps.) Be sure to label each plate with a unique name.

Standard Turn Around Time
Our turnaround time is 8-24 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 should be returned same day and samples received by 4pm should be returned by 10:30 the next business day.