Analysis of blood processing conditions to obtain high-quality total RNA from human leukocyte concentrate.
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.
Pérez
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Genomics and Diagnostic Division, Center for Genetic Engineering and Biotechnology, Havana, Cuba. isabel.guillen@cigb.edu.cu
Santana
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Argudin
T D
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Gardon
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D053607:Adrenomedullin / Q000235:genetics; D001793:Blood Preservation; D002021:Buffers; D020869:Gene Expression Profiling; D006801:Humans; D007962:Leukocytes / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000097:blood*; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction
C507862:ADM protein, human; D002021:Buffers; D012333:RNA, Messenger; D053607:Adrenomedullin; D012313:RNA