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Analysis of blood processing conditions to obtain high-quality total RNA from human leukocyte concentrate
Pérez, I. A; Santana, S. P; Argudin, T. D; Gardon, D. O.
  • Pérez, I. A; Center for Genetic Engineering and Biotechnology. Genomics and Diagnostic Division. Havana. CU
  • Santana, S. P; Center for Genetic Engineering and Biotechnology. Genomics and Diagnostic Division. Havana. CU
  • Argudin, T. D; Center for Genetic Engineering and Biotechnology. Genomics and Diagnostic Division. Havana. CU
  • Gardon, D. O; Center for Genetic Engineering and Biotechnology. Genomics and Diagnostic Division. Havana. CU
Genet. mol. res. (Online) ; 6(2): 298-307, 2007. tab, graf
Article in English | LILACS | ID: lil-482040
ABSTRACT
Blood samples are used as a biological source to discover biomarkers of hematological and non-hematological disorders. The present study shows the impact of different experimental conditions associated with cell lysis buffer, TRI-reagent protocol and blood cell storage buffer and their correlation with the quantity, quality and Adrenomedullin gene expression levels of total RNA when RT-PCR technique is used. A leukocyte cell bank protocol is also proposed for further mRNA expression analysis using RNAlater as storage buffer. There is evidence that total RNA isolated from leukocyte concentrate stored for 1 month at -70 degrees C did not show significant differences concerning quality, purity and Adrenomedullin gene expression compared with the freshly processed leukocyte sample.
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Full text: Available Index: LILACS (Americas) Main subject: Blood Preservation / RNA / RNA, Messenger / Leukocytes Limits: Humans Language: English Journal: Genet. mol. res. (Online) Journal subject: Molecular Biology / Genetics Year: 2007 Type: Article Affiliation country: Cuba Institution/Affiliation country: Center for Genetic Engineering and Biotechnology/CU

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Full text: Available Index: LILACS (Americas) Main subject: Blood Preservation / RNA / RNA, Messenger / Leukocytes Limits: Humans Language: English Journal: Genet. mol. res. (Online) Journal subject: Molecular Biology / Genetics Year: 2007 Type: Article Affiliation country: Cuba Institution/Affiliation country: Center for Genetic Engineering and Biotechnology/CU