Sanger Sequencing FAQs

Basic Information on Sanger Sequencing

  1. Intro to the UM Advanced Genomics Core Sanger Sequencing Service
  2. What types of templates can the Sequencing Core process?
  3. What will I get for my money?
  4. What if my DNA is longer than your machines can read?
  5. Does the Core perform Shotgun sequencing?
  6. What QA/QC measures does the Core implement to minimize mistakes?

How to prepare samples for Sanger sequencing

  1. How to prepare DNA.
  2. Why can’t I use a spectrophotometer to measure my mini-prep DNA?
  3. Can I directly sequence a PCR product?
  4. Can I get sequence from large-insert clones like BACs?
  5. Can I get sequence directly from bacterial genomic DNA?
  6. What primers does the Core provide?
  7. How do I design my own primers?
  8. The correct concentration and volume for your template(s) and primer(s).
  9. How do I label my samples?

Submitting your Sanger samples

  1. Can I Get Rush Processing?
  2. What kind of tubes should I use?
  3. Can I submit my samples in 96-well plates?

After you’ve submitted Sanger samples

  1. I made a mistake entering my sample! How can I correct it?
  2. How long will it take?
  3. How will I get my results back (Sequences and Chromatogram Files)?
  4. My sample is listed as ‘done’, but I can’t find the data!

About your Sanger Sequencing Results

  1. I’ve forgotten my password! How can I get it back so I can retrieve my data?
  2. Can’t I get a hard-copy printout of my chromatogram file?
  3. I downloaded my chromatogram file. Now, how do I open it?
  4. My files are missing from the Download Server! Why???
  5. How long are files kept on the data download server?
  6. How do I interpret the results?
  7. I need some old sequence data from [2013/2016/whenever]. How do I request it?

In Case of Problems

  1. Why won’t the computer let me submit samples?!
  2. My Login has been disabled due to an email problem! Why, and what should I do?
  3. The Results email I received cuts off partway through the message! What should I do?
  4. My files are missing from the Download Server! Why???
  5. Why is my text sequence slightly different than my chromatogram’s sequence?
  6. My primer works fine for PCR – why can’t you sequence with it?
  7. My samples produced only blank lanes. Why?
  8. The technicians reported ‘poor resolution’ for my samples. Why?
  9. Sequencing worked – sort of – the data aren’t very good. Why?
  10. What are the most common reasons for a sequencing reaction to fail?
  11. I think the Core may have made a mistake with my samples.
  12. How do I contact the Core personnel?