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Planarian High Molecular Weight DNA Isolation by Spooling

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Planarian Regeneration

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1774))

Abstract

High-quality large molecular weight genomic DNA is important for genomic studies. Most commercial available genomic DNA purification kits have failed to generate high molecular weight DNA of sufficient quality from planarians. Here, we describe a simple and efficient genomic DNA isolation method, which has worked for several different planarian species, including Schmidtea mediterranea. This phenol–chloroform based method can be used to obtain genomic DNA of up to 150 kb and can be used for bacterial artificial chromosome (BAC) library construction, next-generation sequencing and PCR cloning.

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References

  1. Gregory SG, Sekhon M, Schein J, Zhao S, Osoegawa K, Scott CE et al (2002) A physical map of the mouse genome. Nature 418:743–750

    Article  CAS  Google Scholar 

  2. Osoegawa K, de Jong PJ (2004) BAC library construction. In: Zhao S, Stodolsky M (eds) Bacterial artificial chromosomes volume 1: library construction, physical mapping, and sequencing, 1st edn. Humana Press, Totowa, NJ, p 1

    Google Scholar 

  3. Li R, Fan W, Tian G, Zhu H, He L, Cai J et al (2010) The sequence and de novo assembly of the giant panda genome. Nature 463:311–317

    Article  CAS  Google Scholar 

  4. Wang K, Wang Z, Li F, Ye W, Wang J, Song G et al (2012) The draft genome of a diploid cotton Gossypium raimondii. Nat Genet 44:1098–1103

    Article  CAS  Google Scholar 

  5. Huddleston J, Ranade S, Malig M, Antonacci F, Chaisson M, Hon L et al (2014) Reconstructing complex regions of genomes using long-read sequencing technology. Genome Res 24:688–696

    Article  CAS  Google Scholar 

  6. Voskoboynik A, Neff NF, Sahoo D, Newman AM, Pushkarev D, Koh W et al (2013) The genome sequence of the colonial chordate, Botryllus schlosseri. Elife 2:e00569

    Article  Google Scholar 

  7. Bowen ID, Ryder TA, Winters C (1975) The distribution of oxidizable mucosubstances and polysaccharides in the planarian Polycelis tenuis Iijima. Cell Tissue Res 161:263–275

    Article  CAS  Google Scholar 

  8. Cantarel BL, Korf I, Robb SM, Parra G, Ross E, Moore B et al (2008) MAKER: an easy-to-use annotation pipeline designed for emerging model organism genomes. Genome Res 18:188–196

    Article  CAS  Google Scholar 

  9. Garcia-Fernandez J, Baguna J, Salo E (1993) Genomic organization and expression of the planarian homeobox genes Dth-1 and Dth-2. Development 118:241–253

    CAS  PubMed  Google Scholar 

  10. Cebria F, Newmark PA (2005) Planarian homologs of netrin and netrin receptor are required for proper regeneration of the central nervous system and the maintenance of nervous system architecture. Development 132:3691–3703

    Article  CAS  Google Scholar 

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Correspondence to Alejandro Sánchez Alvarado .

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Zhang, S., Alvarado, A.S. (2018). Planarian High Molecular Weight DNA Isolation by Spooling. In: Rink, J. (eds) Planarian Regeneration. Methods in Molecular Biology, vol 1774. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7802-1_8

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  • DOI: https://doi.org/10.1007/978-1-4939-7802-1_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7800-7

  • Online ISBN: 978-1-4939-7802-1

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