How to Submit Samples for Sequencing:
Sequencing Options
1) Samples Ready for Sequencing: Before submitting ready to sequence samples, please heat-denature at 94 or 95 C in 10-15 µl HiDi Formamide for 3 minutes and place on ice for 5 minutes. Formamide is available at no extra cost from the Core Lab. It is covered under the sequencing fee. If you are unable to add the Formamide and denature samples close (within 4 hours) to the time they will be sequenced you can submit dried down samples after sephadex clean-up (for Sequencing only, not Fragment analysis). The dried down samples are stable for long periods of time in the freezer. You must enter
sample names into the computer if you want them in addition to well ID numbers which are automatic. Ask a Core LabTech for a sample sheet template if you want to enter sample names on your own computer ahead of time. Data files can be picked up in person with a jump drive, CD, or Zip disk or they can be posted to the Core web page and are password protected; please contact the Core Lab for a username and password. For service and fee information follow this link. If you want to know how to load a 96-well plate, run a cycle sequence reaction, or learn about instrument setup options and sequence read lengths follow these links.
2) Clean PCR or Plasmid samples for Cycle Sequencing, Sephadex clean-up, and Sequencing: If you are submitting PCR product and primers for cycle sequencing we recommend a clean-up process using some type of PCR purification kit or protocol or a Plasmid prep such as alkaline lysis prep and ion-exchange resin (such as the columns found in Qiagen's 'REAL' prep or Promega's 'Wizard' prep kits). Following this clean-up we ask that you run a gel with a standard to determine sample concentrations (after clean-up). We want a picture of this gel with each sample band labeled so when we set up the cycle sequence reaction we can make sure the concentrations are high enough for cycle sequencing. If the concentration is less than recommended there may not be any sequence results. The Core Lab techs will label your sequences by sample name and primer. Data files can be picked up in person with a jump drive, CD, or Zip disk or they can be posted to the Core web page and are password protected; please contact the Core Lab for a username and password. For service and fee information follow this link. If you want to learn about instrument setup options and sequence read lengths follow this link.
To process samples for Cycle Sequencing and Sequencing we ask that you provide us with the following:
- Your contact information, Name, Lab name, phone and e-mail
- 25ul of sample (recommended concentration or higher) in a sealed tube, labeled with sample name.
- A picture of a gel after samples were cleaned-up, label bands with sample names and amount of sample loaded in the gel, please include a ladder.
- An easy to understand list of which samples you want run with which primers.
- Primers in labeled tubes with primer name and concentration in pmol/ul or uM solution. We need at least 2ul per sample of 3.2pmol primer. You can give us primers as 3.2 uM, 10 uM, or 100 uM solutions in sufficient volume for the number of samples you need processed, plus extra in case of reruns. ( Note: 1pmol/ul= 1 uM solution)
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