Dilution cloning

Dilution cloning or cloning by limiting dilution {{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}}{{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}} describes a procedure to obtain a monoclonal 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 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, aliquots of the suspension are plated or placed in wells and incubated. If all works correctly, a monoclonal cell colony will be produced. Applications for the procedure include cloning of parasites,{{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 }} T cells,{{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}} transgenic cells,{{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 }} macrophages.{{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 }} and hematopoietic stem cells.{{Cite journal |last1=Sieburg |first1=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 |pmid=12482506 }}