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Agilent RNA Kits for the 2100 Bioanalyzer System Fast quality control of RNA with minimal sample consumption Do you check the quality of your total RNA preparation after isolation? Are you confident about the quality of your mRNA, Cy5 labeled RNA, or guide RNA? Agilent offers RNA assays that allow characterization of a variety of RNA sample types and estimation of their concentrations within minutes, with only a few nano- or picograms of sample. Advantages

  • Minimal sample consumption – use as little as 200 pg of total RNA for analysis, saving most of your valuable sample
  • Faster results – complete automated analysis of 11 or 12 samples in about 30 minutes
  • High assay accuracy and precision – pre-packaged reagents and standardized assay protocols yield highly accurate and reproducible data
  • Quick and easy sample comparison – automated sample alignment, one-click overlay, scaling and zooming features
  • Conveniently archived and stored digital data – easily share data with others and export it for publications or presentations
  • Alternative data display options – results shown in gel-like image, ele- tropherogram, and tabular formats
  • Easy-to-use – simply load the chip, press "start" and the 2100 Bioanalyzer system does the rest
  • RNase free – uses RNase free reagents and chips to avoid sample degradation during analysis
  • Clean – minimal exposure to ethidium bromide or other hazardous materials Analytical specifications RNA 6000 Nano Kit RNA 6000 Pico Kit Small RNA Kit Total RNA Nano mRNA Nano Total RNA Pico mRNA Pico Small RNA Quantitative range 25–500 ng/μL 25–500 ng/μL - - 50–2000 pg/µL of purified miRNA* Qualitative range 5–500 ng/μL 25–250 ng/μL 50–5000 ng/μL* 250–5000 ng/μL* 50–2000 pg/µL of purified miRNA* Quantification reproducibility 10% CV 10% CV 20% CV 20% CV 25% CV Size range - - - - 6–150 nt Sensitivity (S/N>3)* 5 ng/μL 25 ng/μL 50 pg/μL 200 pg/μL (in TE) 250 pg/μL 500 pg/μL (in TE) 50 pg/µL** Quantification accuracy 20% 20% 30% 30% - Maximum sample buffer strength 100 mM Tris,

0.1 EDTA

100 mM Tris, 50 mM Tris, 50 mM Tris, 0.1 mM EDTA Physical specifications Analysis run time 30 minutes 30 minutes 30 minutes 30 minutes 30 minutes Samples per chip 12 12 11 11 11 Sample volume 1 μL 1 μL 1 μL 1 μL 1 μL Kit Stability*** 4 months 4 months 4 months 4 months 4 months *Samples in water Measured for the 40 nt fragment of the Small RNA ladder *Minimum guarantee

[nt] Ladder E1-ZFP42 E2-ZFP42 E3-ZFP42 E1-GUSBE 2-GUSBE 3-GUSBV E1-GUSB VE2-GUSBV E3-GUSBVE1-ZFP42 VE2-ZFP42 VE3-ZFP42 100 150 100 [nt] 200 400 600 800 1,000 1,200 M1-Z M2-Z M3-Z VM1-Z VM2-Z VM3-Z M1-G M2-G M3-G VM1-G VM2-G VM3-G Control E1-Z E2-Z E3-Z VE1-Z VE2-Z VE3-Z E1-G E2-G E3-G VE1-G VE2-G VE3-G Minimal Design ZFP42 Minimal Design GUSB Extended Design ZFP42 Extended Design GUSB Quantity (pg//uni00B5L) Quantitation of Minimal Design Guide RNA (gRNA) RNA 6000 Nano and Pico Assay Example: Quality control of RNA

  • Identify degradation of total RNA
  • Identify degradation of mRNA
  • High sensitivity with minimal sample consumption
  • Unparalleled sensitivity in the picogram range RNase degradation of RNA samples is a common reason for failed experiments. The 2100 Bioanalyzer system provides RNA quality control results in both gel- like image as well as electrophoretic data making it easy to detect even small degradation effects. Indications for RNA degradation are:
  • Decreasing ratio of ribosomal bands
  • Additional peaks below the ribosomal bands
  • Decrease in overall RNA signal
  • Shift towards shorter fragments RNA 6000 Nano and Pico Assay Example: Purity analysis of mRNA
  • Detect and visualize ribosomal RNA contamination in mRNA samples
  • Determine the percentage of ribosomal contamination Many applications, such as cDNA syn- thesis for library construction or RT-PCR profit from highly enriched mRNA. The RNA kits allow the visualization of small impurities of ribosomal RNA in mRNA samples. The percentage calculation of ribosomal contamination and a good estimate of RNA concentration are also shown. All of this can be achieved with minimal amounts of sample. Small RNA Assay Example: Quality control in genome editing
  • Determination of yield and sizing of guide RNAs after in vitro transcription
  • Identification of impurities, truncations, secondary structure formation Small RNA Assay Example: Monitoring of Small RNAs
  • Detect and visualize miRNA directly in total RNA or enriched samples
  • Measure and compare various small RNA regions in different extracts and tissues
  • Monitor and compare expression under various conditions In CRISPR/Cas workflows, the synthesis of guide RNA (gRNA) is a key component for successful genome editing. gRNA created by in vitro transcription and puri- fied may be run on the 2100 Bioanalyzer system. The Small RNA Kit is ideally suited for this application as it allows for high resolution separation of small RNAs from 6-150 nucleotides. Important features to look for: impurities or secondary structure, samples are of expected size and sufficient yield for follow-up studies. Small RNAs sized below 200 nt are hard to separate and measure at low concen - trations. For better results in downstream experiments, e.g. miRNA microarrays or real time PCR, it is beneficial to know what quality and amounts of miRNA are available. www.agilent.com/genomics/bioanalyzer For Research Use Only. Not for use in diagnostic procedures. This information is subject to change without notice. © Agilent Technologies, Inc. 2017 Published in USA, February 1, 2017 5991-7891EN