e Assay Consistency Extraction of DNA or RNA from Blood 2. Extraction of highly concentrated (>100ng/µl) DNA from whole blood samples This article describes a DNA isolation method that does not require the laborious extraction from buffy coat samples that characterizes most approaches to high-concentration (>100ng/µl) genomic DNA extraction. 100 90 80 50 70 60 40 30 20 10 0 Maxwell® 16 LEV Blood DNA Kit (n = 5) QIAamp® Blood Kit (n = 6) Precipitation-based DNA purification protocol (n = 6) Higher DNA recovery from whole blood samples. Genomic DNA was isolated from 200µl of liquid blood using the QIAamp® Blood Kit and from 300µl of liquid blood using the Maxwell® 16 LEV Blood DNA Kit and a precipitation-based DNA purification method. All three sample sets were derived from the same volume of liquid blood on the same day. One standard deviation is indicated for each set. Percent yield was estimated by comparing actual yield to theoretical gDNA yield, based on the white blood cell count of the samples. 10 12 14 16 18 20 0 2 4 6 8 3. Extraction of RNA from 2.5-8ml whole blood samples The Maxwell® 16 LEV simplyRNA Blood Kit is designed for isolation of total RNA from fresh whole blood collected in EDTA tubes. The simplyRNA Blood procedure purifies total RNA with minimal sample handling. The standard protocol can process 2.5ml of fresh whole blood per RNA isolation. In this report, we test the scalability of the simplyRNA blood procedure. 2.5ml blood TM372 5.0ml blood Modified Needle Shearing 8.0ml blood Modified + 8.0ml blood Modified + DNase I RNA was isolated from 2.5, 5 and 8ml of blood using the Maxwell® 16 LEV simplyRNA Blood Kit following the standard protocol outlined in the Maxwell® 16 LEV simplyRNA Blood Kit Technical Manual, and a modified protocol as described here. Try in Your Lab! enews | Fall 2013 % Yield 9071MA Yield (µg) 11125MA Pagina 6
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