RESUMO
OBJECTIVE@#To construct rapidly a full-length cDNA library from nanogram amounts total RNA of Giardia lamblia (G. lamblia) trophozoites stocked in RNA stabilization reagent.@*METHODS@#Total RNA of Giardia was extracted using Trizol reagent. A full-length cDNA library of G. lamblia trophozoites was constructed by a long-distance PCR (LD-PCR) method. The recombinant rate and the coverage rate of full-length clones of the library were evaluated. The inserted fragments were identified and sequenced by PCR amplification.@*RESULTS@#The titer of cDNA library was 3.85 × 10(7) pfu/mL. The length of inserted fragments ranged from 0.4 to 2.5 kb, and the recombination efficiency accounted for 100% (20/20). The coverage rate of full-length clones is high (17/20).@*CONCLUSIONS@#The RNA stabilization reagent may be used to fix the cells and prevent the RNA in cells even though delivered under normal atmospheric temperature. The long-distance PCR can be used to construct a full-length cDNA library rapidly and it needs less RNA than the traditional method from mRNA.
Assuntos
Humanos , DNA de Protozoário , Química , Genética , Biblioteca Gênica , Giardia lamblia , Química , Genética , Giardíase , Parasitologia , Reação em Cadeia da Polimerase , Métodos , RNA , Química , Genética , Trofozoítos , QuímicaRESUMO
This is the case of a 71 year old male who developed multiple myeloma (MM) and chronic myelogenous leukemia (CML) within a two year period. The patient initially presented with osteolytic lesions of the lumbar spine, and following the initial work-up a diagnosis of multiple myeloma with an IgG kappa paraproteinemia was made and appropriate treatment was given. Two years later the patient developed a progressively worsening leukocytosis which was found to be due to Philadelphia Chromosome (Ph1) positive CML. The occurrence in the same patient of two distinct hematologic malignancies suggests a neoplastic transformation of a pluripotent stem cell. A review of the literature appears to support the existence of a relationship between MM and CML as well as a relationship between MM and the myeloproliferative disorders.