Analysis of blood processing conditions to obtain high-quality total RNA from human leukocyte concentrate |
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Authors: | Pérez I A G Santana S P Argudin T D Gardon D O P |
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Institution: | Genomics and Diagnostic Division, Center for Genetic Engineering and Biotechnology, Havana, Cuba. isabel.guillen@cigb.edu.cu |
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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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