Individual Tube Submission Policies

We only accept individual tubes submitted in 2.0ml, self-standing tubes with a 0.5ml interior.

*** Individual tube reactions submitted in any other tube type will not be accepted.***

Here are some examples of acceptable tube types:

• 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

Label the tube with your researcher code and template-primer combination. Be sure to make each name unique and easily identifiable as to ease the sample intake process.

External users: No barcode is required for your tube submission.

Internal Yale users: All tubes must be labeled with barcodes. 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.

Place barcodes vertically on the tubes and label the back with your sample name and researcher code as shown in the provided image.

DO NOT LABEL BARCODES HORIZONTALLY ON TUBES!!!

Review more detailed Barcode Labeling Instructions

In addition to our standard protocol, we have alternative sequencing categories* you may choose (in our dropdown menu on the ordering page): Difficult Template, shRNA, BAC, single-stranded, and Phage/Lambda. Most of these are self-explanatory; however the Difficult Template protocol might be useful in achieving better sequencing results if your sample falls into any of these categories:

• Is GC-Rich: above 60% GC or concentrated GC content in a small section
• Contains secondary structures
• Has a higher melting temperature/harder to denature
• Has certain di- or tri-nucleotide repeats

*These alternative protocols are a higher cost than our standard protocol.

All samples must be submitted premixed, with primer and template in one tube. 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
  • If 100-300bp: 40-50ng
  • If 300-500bp: 40-160ng
  • If 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.

Difficult Templates:

• Mix as above
• Specify as Difficult Template when ordering

shRNA:

• Mix as above
• Specify as shRNA when ordering