View source for Biology:Dilution cloning

'''Dilution cloning or cloning by limiting dilution '''<ref name=Fresh>{{cite book|last=Freshney|first=R. Ian|title=Culture of animal cells : a manual of basic technique and specialized applications|year=2010|publisher=Wiley-Blackwell|location=Hoboken, N.J.|isbn=9780470528129|pages=208–211|edition=6th}}</ref><ref name=Dav>{{cite book|editor-last=Davis|editor-first=John M|title=Animal cell culture|url=https://archive.org/details/animalcellcultur00davi|url-access=limited|year=2011|publisher=Wiley-Blackwell|location=Oxford|isbn=978047066658-6|pages=[https://archive.org/details/animalcellcultur00davi/page/n268 239]–240}}</ref>   describes a procedure to obtain a [[Biology:Monoclonal|monoclonal]] [[Biology:Cell|cell]] population starting from a polyclonal mass of cells.
This is achieved by setting up a series of increasing dilutions of the parent (polyclonal) cell culture. A [[Chemistry:Suspension|suspension]] of the parent cells is made. Appropriate dilutions are then made, depending on cell number in the starting population, as well as the viability and characteristics of the cells being cloned.
After the final dilutions are produced, [[Chemistry:Chemistry|aliquots]] of the suspension are plated<ref name=Fresh />  or placed in wells<ref name=Dav />  and incubated. If all works correctly, a monoclonal cell colony will be produced. Applications for the procedure include cloning of [[Earth:Parasitism|parasite]]s,<ref name="ButterworthRobertson2011">{{cite journal|last1=Butterworth|first1=Alice S|last2=Robertson|first2=Alan J|last3=Ho|first3=Mei-Fong|last4=Gatton|first4=Michelle L|last5=McCarthy|first5=James S|last6=Trenholme|first6=Katharine R|title=An improved method for undertaking limiting dilution assays for in vitro cloning of Plasmodium falciparum parasites|journal=Malaria Journal|volume=10|issue=1|year=2011|pages=95|issn=1475-2875|doi=10.1186/1475-2875-10-95|pmc=3089786|pmid=21496350 |doi-access=free }}</ref> [[Biology:T cell|T cell]]s,<ref>{{cite journal|last=Rout|first=Namita|author2=Else, James G.|author3=Yue, Simon|author4=Connole, Michelle|author5=Exley, Mark A.|author6=Kaur, Amitinder|title=Heterogeneity in phenotype and function of CD8+ and CD4/CD8 double-negative Natural Killer T cell subsets in sooty mangabeys|journal=Journal of Medical Primatology|date=2 July 2010|volume=39|issue=4|pages=224–234|doi=10.1111/j.1600-0684.2010.00431.x|pmc=2904642|pmid=20618588}}</ref> transgenic cells,<ref>{{cite journal|last=Lilienfeld|first=Benjamin G.|author2=Crew, Mark D.|author3=Forte, Pietro|author4=Baumann, Bettina C.|author5=Seebach, Jörg D.|title=Transgenic expression of HLA-E single chain trimer protects porcine endothelial cells against human natural killer cell-mediated cytotoxicity|journal=Xenotransplantation|date=1 March 2007|volume=14|issue=2|pages=126–134|doi=10.1111/j.1399-3089.2007.00378.x|pmid=17381687|s2cid=1433274 }}</ref> [[Biology:Macrophage|macrophage]]s.<ref>{{cite journal|last=Seki|first=Yoshihiro|author2=Suzuki, Satoshi O.|author3=Masui, Kenta|author4=Harada, Shiori|author5=Nakamura, Seiji|author6=Kanba, Shigenobu|author7=Iwaki, Toru|title=A simple and high-yield method for preparation of rat microglial cultures utilizing Aclar plastic film|journal=Neuropathology|date=1 June 2011|volume=31|issue=3|pages=215–222|doi=10.1111/j.1440-1789.2010.01163.x|pmid=21092060|s2cid=26016732 }}</ref> and [[Biology:Hematopoietic stem cell|hematopoietic stem cells]].<ref>{{Cite journal |last=Sieburg |first=Hans B |last2=Cho |first2=Rebecca H |last3=Muller-Sieburg |first3=Christa E |date=2002 |title=Limiting dilution analysis for estimating the frequency of hematopoietic stem cells: uncertainty and significance |url=https://www.exphem.org/article/S0301-472X(02)00963-3/fulltext |journal=Exp Hematol |volume=39 |issue=12 |pages=1436–1443 |doi=10.1016/s0301-472x(02)00963-3 |via=ISEH|doi-access=free }}</ref>

== References ==
{{Reflist}}

==External links==
*Robert Staszewski "Cloning by Limiting Dilution: an improved estimate that an interesting culture is monoclonal" [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2589791/]
*John A. Ryan [https://www.corning.com/catalog/cls/documents/protocols/Single_cell_cloning_protocol.pdf "Cell Cloning by Serial Dilution in 96 Well Plates".]
*Vilma Maldonado,Jorge Meléndez-Zajgla [http://www.medigraphic.com/pdfs/bioquimia/bq-2007/bq072e.pdf "A modified method for cloning adherent mammalian cells".]
*[http://people.rit.edu/~gtfsbi/hytc/PDFfiles/Cloning%20Lab.pdf "Cloning by Limiting Dilution".]{{Dead link|date=July 2019 |bot=InternetArchiveBot |fix-attempted=yes }}
*[http://www.sciencegateway.org/protocols/cellbio/cell/cbld.htm "Cloning by limiting dilution".]
*Nanci Donacki [http://www.protocol-online.org/prot/Protocols/Cloning-by-Limiting-Dilution-of-Hybridoma-86.html "Cloning by Limiting Dilution of Hybridoma".]

{{DEFAULTSORT:Dilution Cloning}}
[[Category:Molecular biology techniques|Cloning]]

{{Sourceattribution|Dilution cloning}}