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Individual Tube Submission Policies

Plasticware and Labeling

  1. All samples must be submitted with primer and template premixed.
  2. We only accept individual tubes submitted in 2.0ml, self-standing tubes with a 0.5ml interior. Some examples below:
- DOT Scientific screw cap tubes. Catalog numbers 914-NC (tubes) and 503-SC (caps).
- American Bioanalytical purple screw cap tubes, available from the Stockroom through SciQuest. The catalog number is PL73310-00001 and they are packaged 500 tubes with caps.
- VWR Labcon available through SciQuest, Part #89004-284
- USA Scientific available through SciQuest, Part# 1405-9700, 1405-9701, 1405-9702, 1405-9703, 1405-9704, 1405-9705, 1405-9706, 1405-9799
- Genesee Scientific: catalog #21-258

*** Individual tube premixed reaction submitted in any other tube types will not be accepted.***

3. Internal Yale users: All tubes must be labeled with barcodes , template name and researcher code. Barcodes are given out over the counter in the Kline and Medical Stockrooms, outside of S340 TAC, BCMM 108, 300 George Street, Room 2127 (come visit us!) at no additional cost.

4. External users: Tubes will be barcoded upon receipt. Label the tube with template name and your researcher code.

***Please see Barcode Labeling Instructions for more information regarding proper barcode placement and tube labeling instructions.***

Template preparation

Provide primer and template premixed according to the guidelines below. We offer a high-volume service for large projects. (Plate submissions can contain any number of wells, but there is a 48 well minimum charge.) Please see High Volume Submission Policies and Procedures [ST1] for more information.

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

***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.***

***Nanodrop help guide***

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.

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.