Small-Scale Isolation of High Molecular Weight DNA from Leishmania braziliensis

E. Carmelo; J. A. Pérez; A. I. Zurita; J. Piñero; F. de Armas; A. del Castillo; B. Valladares

doi:10.1645/0022-3395(2000)086[0844:SSIOHM]2.0.CO;2

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E. Carmelo,¹ J. A. Pérez,¹ A. I. Zurita,¹ J. Piñero,¹ F. de Armas,¹ A. del Castillo,¹ B. Valladares²

¹Departamento de Parasitología, Ecología y Genética, Facultad de Farmacia, Universidad de La Laguna, Astrofísico Fco. Sánchez, s/n, 38271 La Laguna, S/C de Tenerife, Spain
²author for correspondence

Abstract

In this paper, we report a method for isolation of high molecular weight DNA from Leishmania promastigotes. This technique is especially indicated for small-scale purification of DNA suitable for the construction of highly representative genomic libraries. In our protocol, lysis buffer is compatible with RNase treatment, avoiding an additional precipitation step and consequent shearing of DNA. In order to prove the quality of the DNA isolated by this method, a Leishmania braziliensis genomic library was constructed, and an L. braziliensis KMP-11 gene was cloned after screening the library with a heterologous probe.

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E. Carmelo, J. A. Pérez, A. I. Zurita, J. Piñero, F. de Armas, A. del Castillo, and B. Valladares "Small-Scale Isolation of High Molecular Weight DNA from Leishmania braziliensis," Journal of Parasitology 86(4), 844-846, (1 August 2000). https://doi.org/10.1645/0022-3395(2000)086[0844:SSIOHM]2.0.CO;2

Published: 1 August 2000

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