NextGen Sequencing - Sample Requirements
DNA
- DNA should be provided as double-stranded high molecular weight DNA with an OD 260/A280 ≥1.8 and an OD 260/230 ≥ 1.9.
- DNA should preferably be dissolved in RNase-, DNase- and protease-free high molecular grade water.
- Samples should be treated with RNase (e.g. from QIAGEN) to minimize contamination through RNA.
- “Ready to load” libraries, PCR products and amplicons must be column purified from low molecular weight impurities (like e.g., primers, and nucleotides) and reaction buffer and should appear as single band on an agarose gel.
RNA
- RNA should be provided as high quality RNA with an OD 260/280 ≥ 1.8 and an OD 260/230 ≥ 1.8.
- The RNA Integrity Number (RIN; Agilent Technologies 2100 Bioanalyzer) resp. RNA quality indicator (RQI; Bio-Rad’s Experion) value should be greater than or equal to 8.
- RNA should preferably be dissolved in RNase-, DNase- and protease-free high molecular grade water or Tris buffer (do not use DEPC-treated H2O).
- We strongly recommend performing a final clean-up of the RNA using commercial available RNA purification kits (e.g. RNeasy spin columns from QIAGEN).
For information on the DNA / RNA quantity and concentration required for specific library preparation methods please refer to your quote or contact us.
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