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High Throughput RNA Isolation for High Performance Microarray Analysis

gh quality and yield of total RNA is critical for successful gene expression profiling experiments. RNA quality is assessed by the A260/A280 ratio, the ratio of 28S:18S rRNA peaks, and RNA Integrity Number (RIN). As shown in Figure 1, the MagMAX-96 for Microarrays Spin and No-Spin Procedures both purify RNA with good yield and quality. RIN is based on the entire electrophoretic trace of the RNA sample (1=highly degraded and 10=highly intact). Total RNA purified using MagMAX-96 for Microarrays has high integrity (RIN typically >7.8).

Figure 1. Obtain RNA of High Quality and Yield with MagMAX-96 for Microarrays Total RNA Isolation Kit. RNA was isolated from frozen mouse liver with the MagMAX-96 for Microarrays Kit. Tissue homogenates were processed in quadruplets or octuplets using the Spin procedure (homogenate derived from 10 mg tissue per sample) or the No-Spin procedure (homogenate derived from 5 mg tissue per sample), respectively. Purified RNA (2 l) was quantified using a NanoDrop Spectrophotometer (A, B, and C). The ratio of 28S to 18S rRNA was determined with an RNA LabChip Kit and the Agilent 2100 bioanalyzer.


Successful Microarray Experiments

Total RNA isolated from mouse kidney using MagMAX-96 for Microarrays or the Affymetrix recommended protocol (Protocol A: phenol-based sample lysis followed by glass fiber filter RNA purification) were compared by microarray analysis. Normalized microarray signal intensities obtained from two total RNA samples isolated using MagMAX-96 for Microarrays (Figure 2A) and from total RNA samples purified using MagMAX-96 for Microarrays and Protocol A (Figur
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