tinely isolate 80-100 yg of RNA per 8 mL of culture. RNA isolated by this method or with TRIzol (manufacturer's recommended procedure) exhibited an A260-280 ratio of 1.6-1.8. When needed, RNA samples were supplemented with RNA loading dye [0.72 mL formamide, 0.16 mL lox MOPS buffer, 0.26 mL 37% formaldehyde, 0.18 mL distilled water (DEPC-treated), 0.10 mL 80% glycerol, and 8.0 mg bromophenol bl...

When optimal conditions are employed, real-time polymerase chain reaction (PCR) is a sensitive and accurate technique enabling the quantiWcation of low-copynumber transcripts [1]. However, as with conventional PCR, small variations in initial reaction conditions are ampliWed exponentially and can signiWcantly aVect results [2]. Uniform reaction conditions are therefore essential to achieve accu...

BACKGROUND HPV oncogenes mRNA detection gains momentum as an adjuvant for HPV-related cervical abnormalities diagnosis, but is based on costly detection assays not allowing viral type targeting. OBJECTIVE To assess detection rate of HPV oncogenes E6/E7 mRNA from cervical specimens using a manual, open technology, fully customizable protocol and determine whether HPV-related epidemiological fe...

There are increasing reports of plasma miRNAs as biomarkers of human disease but few standards in methodologic reporting, leading to inconsistent data. We systematically reviewed plasma miRNA studies published between July 2013-June 2014 to assess methodology. Six parameters were investigated: time to plasma extraction, methods of RNA extraction, type of miRNA, quantification, cycle threshold (...

BACKGROUND AND OBJECTIVES Accurate quantification of BCR-ABL mRNA is of critical importance for managing patients with chronic myeloid leukemia (CML) who are receiving imatinib therapy. RNA degradation thus constitutes a potential problem for laboratories quantifying minimal residual disease (MRD). Patients' samples that take a long time to be transported from the hospital to the analyzing labo...

We read the article by Cook et al. (3) with interest, since they compared their findings with our previous work (4–8; D. Brashear, S. Taylor, J. Xiang, D. Klinzman, F. LaBrecque, M. Pfaller, B. Alden, D. LaBrecque, J. Stapleton, and W. Schmidt, 10th Triennial Int. Symp. Viral Hepatitis Liver Dis., abstr. C008, 2000; J. Xiang, F. LaBrecque, W. N. Schmidt, D. Klinzman, D. Brashear, D. R. LaBrecqu...

We read the article by Cook et al. (3) with interest, since they compared their findings with our previous work (4–8; D. Brashear, S. Taylor, J. Xiang, D. Klinzman, F. LaBrecque, M. Pfaller, B. Alden, D. LaBrecque, J. Stapleton, and W. Schmidt, 10th Triennial Int. Symp. Viral Hepatitis Liver Dis., abstr. C008, 2000; J. Xiang, F. LaBrecque, W. N. Schmidt, D. Klinzman, D. Brashear, D. R. LaBrecqu...

Evaluation of gene expression in urinary sediment has been considered as a promising non-invasive approach for biomarker identification of kidney diseases. Nonetheless, there are several challenges in extraction of RNA from this valuable source of biomarkers, mostly because of the factors that have influence on quality of isolated RNA such as low cellular content. Accordingly, we compared the q...

objective: the current methods for bladder cancer diagnosis suffer from low sensitivity and specificity. therefore, finding a novel tumor markers with high specificity and sensitivity is of great interest. micrornas (mirna, mir) are small endogenously-produced, non-coding rnas with an important role in regulating gene expression. recent studies show that mirnas expression profiles represent sig...

Automated nucleic acid extraction is an attractive alternative to labor-intensive manual methods. We compared two automated methods, the BioRobot M48 instrument (Qiagen, Inc.) and MagNA Pure (Roche Applied Sciences) methods, to two manual methods, the QIAamp Viral RNA Mini kit (Qiagen) and TRIzol (Invitrogen), for the extraction of enterovirus RNA. Analytical sensitivity was assessed by dilutio...