Skip to main content
SpringerLink
Log in

Synchronization of Yeast

  • Protocol
  • First Online:
Cell Cycle Synchronization

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1524))

Abstract

The budding yeast Saccharomyces cerevisiae and fission yeast Schizosaccharomyces pombe are amongst the simplest and most powerful model systems for studying the genetics of cell cycle control. Because yeast grows very rapidly in a simple and economical media, large numbers of cells can easily be obtained for genetic, molecular, and biochemical studies of the cell cycle. The use of synchronized cultures greatly aids in the ease and interpretation of cell cycle studies. In principle, there are two general methods for obtaining synchronized yeast populations. Block-and-release methods can be used to induce cell cycle synchrony. Alternatively, centrifugal elutriation can be used to select synchronous populations. Because each method has innate advantages and disadvantages, the use of multiple approaches helps in generalizing results. An overview of the most commonly used methods to generate synchronized yeast cultures is presented along with working Notes: a section that includes practical comments, experimental considerations and observations, and hints regarding the pros and cons innate to each approach.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
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

References

  1. Forsburg SL (2003) Overview of Schizosaccharomyces pombe. Curr Protoc Mol Biol Chapter 13:Unit 13.14

    PubMed  Google Scholar 

  2. Forsburg SL (2005) The yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe: models for cell biology research. Gravit Space Biol Bull 18(2):3–9

    PubMed  Google Scholar 

  3. Sherman F (2002) Getting started with yeast. Methods Enzymol 350:3–41

    Article  CAS  PubMed  Google Scholar 

  4. Forsburg SL, Rhind N (2006) Basic methods for fission yeast. Yeast 23(3):173–183

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Broach JR, Pringle JR, Jones EW (1991) The Molecular and cellular biology of the yeast Saccharomyces. Cold Spring Harbor monograph series, vol 21. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

    Google Scholar 

  6. Guthrie C, Fink GR (1991) Guide to yeast genetics and molecular biology, vol 194. Methods Enzymol Academic Press, Inc, Pasadena, CA

    Book  Google Scholar 

  7. Murray AW, Hunt T (1993) The cell cycle: an introduction. W.H. Freeman, New York

    Google Scholar 

  8. Jorgensen P, Tyers M (2004) How cells coordinate growth and division. Curr Biol 14:R1014–R1027

    Article  CAS  PubMed  Google Scholar 

  9. Schneider BL, Zhang J, Markwardt J, Tokiwa G, Volpe T, Honey S, Futcher B (2004) Growth rate and cell size modulate the synthesis of, and requirement for, G1-phase cyclins at start. Mol Cell Biol 24:10802–10813

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Fantes P, Brooks R (1993) The cell cycle: a practical approach. The practical approach series. IRL Press at Oxford University Press, Oxford, New York

    Google Scholar 

  11. Johnston JR (1994) Molecular genetics of yeast: a practical approach. The practical approach series, vol 141. IRL Press at Oxford University Press, Oxford, New York

    Google Scholar 

  12. Kume K (2016) Elutriation for cell cycle synchronization in fission yeast. Methods Mol Biol 1342:149–155

    Article  PubMed  Google Scholar 

  13. Tormos-Perez M, Perez-Hidalgo L, Moreno S (2016) Fission yeast cell cycle synchronization methods. Methods Mol Biol 1369:293–308

    Article  PubMed  Google Scholar 

  14. Gomez EB, Forsburg SL (2004) Analysis of the fission yeast Schizosaccharomyces pombe cell cycle. Methods Mol Biol 241:93–111

    PubMed  Google Scholar 

  15. Green MD, Sabatinos SA, Forsburg SL (2009) Microscopy techniques to examine DNA replication in fission yeast. Methods Mol Biol 521:463–482

    Article  CAS  PubMed  Google Scholar 

  16. Humphrey T, Brooks G (2005) Cell cycle control: mechanisms and protocols. Methods in molecular biology, vol 296. Humana Press, Totowa, NJ, p 402

    Google Scholar 

  17. Day A, Schneider C, Schneider BL (2004) Yeast Cell Synchronization. Cell Cycle Checkpoint Control Protocols 241:55–76

    Google Scholar 

  18. Luche DD, Forsburg SL (2009) Cell-cycle synchrony for analysis of S. pombe DNA replication. Methods Mol Biol 521:437–448

    Article  CAS  PubMed  Google Scholar 

  19. Spellman PT, Sherlock G, Zhang MQ, Iyer VR, Anders K, Eisen MB, Brown PO, Botstein D, Futcher B (1998) Comprehensive identification of cell cycle-regulated genes of the yeast Saccharomyces cerevisiae by microarray hybridization. Mol Biol Cell 9:3273–3297

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Futcher B (1999) Cell cycle synchronization. Methods Cell Sci 21:79–86

    Article  CAS  PubMed  Google Scholar 

  21. Walker GM (1999) Synchronization of yeast cell populations. Methods Cell Sci 21:87–93

    Article  CAS  PubMed  Google Scholar 

  22. Johnston LH, Johnson AL (1997) Elutriation of budding yeast. Methods Enzymol 283:342–350

    Article  CAS  PubMed  Google Scholar 

  23. Day A, Schneider C, Schneider BL (2004) Yeast cell synchronization. Methods Mol Biol 241:55–76

    PubMed  Google Scholar 

  24. Sonoda E (2006) Synchronization of cells. Subcel Biochem 40:415–418

    Google Scholar 

  25. Amon A (2002) Synchronization procedures. Methods Enzymol 351:457–467

    Article  CAS  PubMed  Google Scholar 

  26. Marbouty M, Ermont C, Dujon B, Richard GF, Koszul R (2014) Purification of G1 daughter cells from different Saccharomycetes species through an optimized centrifugal elutriation procedure. Yeast 31:159–166

    Article  CAS  PubMed  Google Scholar 

  27. Wang S, Luo C (2016) Cell cycle synchronization using a microfluidic synchronizer for fission yeast cells. Methods Mol Biol 1342:259–268

    Article  PubMed  Google Scholar 

  28. Tian Y, Luo C, Lu Y, Tang C, Ouyang Q (2012) Cell cycle synchronization by nutrient modulation. Integr Biol (Camb) 4:328–334

    Article  CAS  Google Scholar 

  29. Hur JY, Park MC, Suh KY, Park SH (2011) Synchronization of cell cycle of Saccharomyces cerevisiae by using a cell chip platform. Mol Cells 32:483–488

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Yang J, Dungrawala H, Hua H, Manukyan A, Abraham L, Lane W, Mead H, Wright J, Schneider BL (2011) Cell size and growth rate are major determinants of replicative lifespan. Cell Cycle 10:144–155

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Bean JM, Siggia ED, Cross FR (2006) Coherence and timing of cell cycle start examined at single-cell resolution. Mol Cell 21:3–14

    Article  CAS  PubMed  Google Scholar 

  32. Di Talia S, Skotheim JM, Bean JM, Siggia ED, Cross FR (2007) The effects of molecular noise and size control on variability in the budding yeast cell cycle. Nature 448:947–951

    Article  PubMed  Google Scholar 

  33. Schmoller KM, Turner JJ, Koivomagi M, Skotheim JM (2015) Dilution of the cell cycle inhibitor Whi5 controls budding-yeast cell size. Nature 526:268–272

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Amberg DC, Burke D, Strathern JN (2005) Methods in yeast genetics : a Cold Spring Harbor Laboratory course manual, 2005th edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

    Google Scholar 

  35. Hasek J (2006) Yeast fluorescence microscopy. Methods Mol Biol 313:85–96

    CAS  PubMed  Google Scholar 

  36. Banfalvi G (2008) Cell cycle synchronization of animal cells and nuclei by centrifugal elutriation. Nat Protoc 3:663–673

    Article  CAS  PubMed  Google Scholar 

  37. Banfalvi G (2011) Synchronization of mammalian cells and nuclei by centrifugal elutriation. Methods Mol Biol 761:25–45

    Article  CAS  PubMed  Google Scholar 

  38. Manukyan A, Abraham L, Dungrawala H, Schneider BL (2011) Methods Mol Biol 761:173–199

    Article  CAS  PubMed  Google Scholar 

  39. Ausubel FM (1987) Current protocols in molecular biology. Greene Publishing Associates, Brooklyn, N. Y & Media, Pa

    Google Scholar 

  40. Futcher B (2002) Transcriptional regulatory networks and the yeast cell cycle. Curr Opin Cell Biol 14:676–683

    Article  CAS  PubMed  Google Scholar 

  41. Chu S, DeRisi J, Eisen M, Mulholland J, Botstein D, Brown PO, Herskowitz I (1998) The transcriptional program of sporulation in budding yeast. Science 282:699–705

    Article  CAS  PubMed  Google Scholar 

  42. Primig M, Williams RM, Winzeler EA, Tevzadze GG, Conway AR, Hwang SY, Davis RW, Esposito RE (2000) The core meiotic transcriptome in budding yeasts. Nat Genet 26:415–423

    Article  CAS  PubMed  Google Scholar 

  43. Haase SB, Reed SI (2002) Improved flow cytometric analysis of the budding yeast cell cycle. Cell Cycle 1:132–136

    Article  CAS  PubMed  Google Scholar 

  44. Sabatinos SA, Forsburg SL (2009) Measuring DNA content by flow cytometry in fission yeast. Methods Mol Biol 521:449–461

    Article  CAS  PubMed  Google Scholar 

  45. Zhang H, Siede W (2004) Analysis of the budding yeast Saccharomyces cerevisiae cell cycle by morphological criteria and flow cytometry. Methods Mol Biol 241:77–91

    PubMed  Google Scholar 

  46. Breeden LL (1997) Alpha-factor synchronization of budding yeast. Methods Enzymol 283:332–341

    Article  CAS  PubMed  Google Scholar 

  47. Richardson HE, Wittenberg C, Cross F, Reed SI (1989) An essential G1 function for cyclin-like proteins in yeast. Cell 59:1127–1133

    Article  CAS  PubMed  Google Scholar 

  48. Schneider BL, Patton EE, Lanker S, Mendenhall MD, Wittenberg C, Futcher B, Tyers M (1998) Yeast G1 cyclins are unstable in G1 phase. Nature 395:86–89

    Article  CAS  PubMed  Google Scholar 

  49. Woldringh CL, Fluiter K, Huls PG (1995) Production of senescent cells of Saccharomyces cerevisiae by centrifugal elutriation. Yeast 11:361–369

    Article  CAS  PubMed  Google Scholar 

  50. Egilmez NK, Chen JB, Jazwinski SM (1990) Preparation and partial characterization of old yeast cells. J Gerontol 45:B9–B17

    Article  CAS  PubMed  Google Scholar 

  51. Day A, Markwardt J, Delaguila R, Zhang J, Purnapatre K, Honigberg SM, Schneider BL (2004) Cell size and Cln-Cdc28 complexes mediate entry into meiosis by modulating cell growth. Cell Cycle 3:1433–1439

    Article  CAS  PubMed  Google Scholar 

  52. Nachman I, Regev A, Ramanathan S (2007) Dissecting timing variability in yeast meiosis. Cell 131:544–556

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Cell Biology and Biochemistry, Texas Tech University Health Sciences Center, Lubbock, TX, USA

    Jessica Smith

  2. Department of Biochemistry and Molecular Genetics, University of Virginia, School of Medicine, Charlottesville, VA, USA

    Arkadi Manukyan

  3. Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, USA

    Hui Hua & Huzefa Dungrawala

  4. Department of Medical Education, Texas Tech University Health Sciences Center, 3601 4th St Rm 5C119, Lubbock, TX, 79430, USA

    Brandt L. Schneider

Authors
  1. Jessica Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Arkadi Manukyan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hui Hua
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Huzefa Dungrawala
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Brandt L. Schneider
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Brandt L. Schneider .

Editor information

Editors and Affiliations

  1. Department of Biotechnology and Microbiology, University of Debrecen, Debrecen, Hungary

    Gaspar Banfalvi

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media New York

About this protocol

Cite this protocol

Smith, J., Manukyan, A., Hua, H., Dungrawala, H., Schneider, B.L. (2017). Synchronization of Yeast. In: Banfalvi, G. (eds) Cell Cycle Synchronization. Methods in Molecular Biology, vol 1524. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6603-5_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-6603-5_14

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6602-8

  • Online ISBN: 978-1-4939-6603-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation