Small-scale isolation of high molecular weight DNA from Leishmania braziliensis

In this paper, we report a method for isolation of high molecular weight DN A from Leishmania promastigotes. This technique is especially indicated for small-scale purification of DNA suitable for the construction of highly re presentative genomic libraries. In our protocol, lysis buffer is compatible with RNase treatment. avoiding an additional precipitation step and conseq uent 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.