Small-scale isolation of high molecular weight DNA from Leishmania braziliensis.
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.
Carmelo
E
E
Departamento de Parasitología, Ecología y Genética, Facultad de Farmacia, Universidad de La Laguna, La Laguna, S/C de Tenerife, Spain.
Pérez
J A
JA
Zurita
A I
AI
Piñero
J
J
de Armas
F
F
del Castillo
A
A
Valladares
B
B
D000818:Animals; D010582:Bacteriophage lambda; D016054:DNA, Protozoan / Q000737:chemistry / Q000302:isolation & purification* / Q000592:standards; D015723:Gene Library; D007892:Leishmania braziliensis / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015800:Protozoan Proteins / Q000235:genetics; D015183:Restriction Mapping
D016054:DNA, Protozoan; D008562:Membrane Glycoproteins; D015800:Protozoan Proteins; C090009:kinetoplastid membrane protein 11, Leishmania donovani