AllPrep DNA/RNA Micro Kit

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RNA isolation kits comparison: RNA isolation kits were compared with each other and with kits for parallel isolation. Comparison of kits was performed on buffy coat samples with defined numbers of cells in the follow- ing aliquots: 1, 10, 20, 50, 100, 150 and 200×103 cells sorted by FACS. Aliquots were defined based on previous experience with availability of target cell population in MGs. Samples were tested in duplicate. The average yield from each aliquot is shown in figure 2. RNA concentrations and integrity numbers (RINs) were evaluated using RNA 6000 Pico kit on Bioanalyzer2100 (Agilent Technologies). The concentration can be also measured with RNA HS assay kit on Qubit 2.0 fluorom- eter (Thermo Fisher Scientific) but this measurement does not allow evaluation of RIN number and we did not use it in our comparisons.Of all of the kits, the NucleoSpin TriPrep (Macherey-Nagel) showed the highest yield but the concentration per microlitre of elution buffer was very low in comparison with AllPrepDNA/ RNA Micro Kit (Qiagen) and RNeasy Micro Kit (Qiagen). The main reason was an almost three times higher volume of elution buffer: 40 mL for NucleoSpin TriPrep (Macherey-Nagel) versus 14 mL for AllPrep DNA/RNA Micro Kit and RNeasy Micro Kit (Qiagen). Both magnetic kits (MagMAX Total RNA Isolation Kit and μMACS mRNA Isolation Kit) were not sufficient for successful isolation of RNA from small amounts of input mater- ial (figure 2).

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