Skip to main content

Quantitation of DNA-Encoded Libraries by qPCR

  • Protocol
  • First Online:
DNA-Encoded Chemical Libraries

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2541))

  • 1238 Accesses

Abstract

DNA-encoded library (DEL) yields can be easily measured throughout the selection process using the quantitative polymerase chain reaction (qPCR) (Sannino A, Gabriele E, Bigatti M, Mulatto S, Piazzi J, Scheuermann J, Neri D, Donckele EJ, Samain F, Chembiochem Eur J Chem Biol 20:955–962, 2019). Samples taken throughout the selection process are diluted prior to amplification and compared to standards of known DNA concentration. Here, I describe a general protocol using a double-stranded DNA binding dye for reaction monitoring. This allows the selection process to be assessed at each step prior to preparation for sequencing. The same method has additional applications in the practice of DEL technology.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. VanGuilder HD, Vrana KE, Freeman WM (2008) Twenty-five years of quantitative PCR for gene expression analysis. BioTechniques 44:619–626. https://doi.org/10.2144/000112776

    Article  CAS  PubMed  Google Scholar 

  2. Taylor SC, Nadeau K, Abbasi M, Lachance C, Nguyen M, Fenrich J (2019) The ultimate qPCR experiment: producing publication quality, reproducible data the first time. Trends Biotechnol 37:761–774. https://doi.org/10.1016/j.tibtech.2018.12.002

    Article  CAS  PubMed  Google Scholar 

  3. Sannino A, Gabriele E, Bigatti M, Mulatto S, Piazzi J, Scheuermann J, Neri D, Donckele EJ, Samain F (2019) Quantitative assessment of affinity selection performance by using DNA-encoded chemical libraries. Chembiochem Eur J Chem Biol 20:955–962. https://doi.org/10.1002/cbic.201800766

    Article  CAS  Google Scholar 

  4. Andrade H, Thomas AK, Lin W, Reddavide FV, Zhang Y (2020) Using a PCR-based method to analyze and model large, heterogeneous populations of DNA. Chembiochem 21:1144–1149. https://doi.org/10.1002/cbic.201900603

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Christopher S. Kollmann .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2022 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Kollmann, C.S. (2022). Quantitation of DNA-Encoded Libraries by qPCR. In: Israel, D., Ding, Y. (eds) DNA-Encoded Chemical Libraries. Methods in Molecular Biology, vol 2541. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2545-3_17

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-2545-3_17

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2544-6

  • Online ISBN: 978-1-0716-2545-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics