Sample Preparation and Isolation of RNA or DNA
High quality starting material is vital to the success of any RNA or DNA analysis. Asuragen’s extensive experience in a wide range of tissue types and formats, qualified and miRNA-compatible methods, and focus on quality to ensure your studies start with the best RNA or DNA possible.
All our workflows feature process controls, certainty of extensive operator qualification, and our Data Quality Guarantee.
Services Available
Method and Staff Qualification
Quality Control
Sample Handling and Processing
Turn-around Time
Additional Services Available
Requirements for Sample Prep Services
Services Available
Eukaryotic or prokaryotic cell linesBlood and other biofluids
Frozen/RNALater®/RNARetain® tissues
Fixed (FFPE) tissues
Methods may utilize exogenous DNA or RNA spiked-in controls.
Method and Staff Qualification
The Asuragen team has been isolating high-quality RNA and DNA since 2001, with a long history of nucleic acid isolation from diverse human and animal tissue types. Any new isolation protocol or new operator at Asuragen Services must demonstrate quality performance based on objective, quantitative measures before either is put in service for client GLP projects. In the case of isolation protocols, Asuragen measures the following parameters against established benchmarks prior to release of a new GLP protocol:
- Yield – including miRNA fraction
- Quality
- Integrity
- Stability
- Contaminants
When a new team member joins the isolations staff, they are provided hands-on training with Asuragen-provided test materials. After their training period, the quality, purity, and integrity of a qualification set of samples is evaluated on each operator before they are considered “qualified” and allowed to extract RNA or DNA from a client sample. Operator metrics are tracked daily, with appropriate contingencies in the event QC metrics fall below specified levels. We are proud of our team members and they are proud of the work they do on your studies.
Quality Control
No matter how robust the downstream processes, good data is dependent on sample quality; consequently, accurate measurement of sample quality is essential before deciding if a sample merits further analysis. All samples prepared by Asuragen Services are routinely qualified by spectrophotometry with the NanoDrop spectrophotometer and the Agilent 2100 Bioanalyzer. Due to the degraded nature of RNA from FFPE tissues, special quality control measurements are used in place of the Bioanalyzer (see Sample Prep: FFPE). Regardless of downstream application, detailed QC reports including nucleic acid concentration, RNA integrity (RIN) electropherograms (where applicable), and purity are provided on each sample. For FFPE samples, concentration and QC pass/fail information is provided. If you have chosen to use Asuragen Services for additional services and a sample does not pass the recommended QC metrics for analysis, you will be contacted promptly to discuss possible solutions. These may include replacing the sample or re-isolating the RNA, excluding the sample from further analysis, or proceeding at-risk.
Yield:
RNA yield is primarily dependent on the tissue type and mass (or volume) but is often affected by disease status, chemical treatment, media, preservatives, and tissue handling.
Integrity:
RNA integrity is dependent on a wide range of factors in the precise handling of biological materials from collection to RNA isolation. Elimination of the activity of endogenous endo- or exo-nucleases is vital to preserving RNA integrity, and typically depends on treatment with proper preservative or freezing upon tissue collection.
Purity:
RNA purity is dependent on the quality of the RNA isolation method used, the tissue type, fixation methods, and sample degradation.
Variation:
Whether from an “experimental” or “normal” population, there is naturally wide variation in yield, degradation mechanisms, and contaminants. Asuragen has performed RNA isolation from thousands of samples, hundreds of tissue types, and dozens of different formats, and we practice methods optimized and tested for maximum yield, integrity, and purity, but we cannot guarantee the yield, integrity, or purity from any given experimental sample.
Asuragen’s rigorous procedures enable Asuragen to be the only service provider to offer a Data Quality Guarantee:
- If, for any reason, a qualified sample does not pass our process control specifications, we will re-run the sample at no additional charge.
Read more about our committment to the best possible data quality, or read about our contributions to in-process controls and standardization.
Sample Handling and Processing
All samples are received and entered into the isolation queue under a controlled SOP to confirm package/sample integrity, appropriate shipping conditions (frozen samples/dry ice, as appropriate), and individual confirmation of sample identity against the shipping manifest and electronic sample submission documents.
Standard RNA isolation quality control criteria include yield, purity, and integrity. All Asuragen protocols are qualified by these criteria in addition to RNA stability, endonuclease and exonuclease contamination, and residual DNA contamination measurements. These additional QC measures are also available for your RNA isolations at an additional charge. Unless noted, all RNA isolation methods are optimized to recover miRNA.
Typical yield, purity, integrity, stability, DNA contamination, and endo/exonuclease data on a wide variety of tissues and cell lines are available for inspection.
Turn-around Time
Most projects are completed in two weeks or less. If you have a critical time requirement, we can often meet your need for no additional charge. Read more about our commitment to prompt turn-around times.
Additional Services Available
mRNA Analysis:
Affymetrix Gene Expression Profiling Service
Illumina Gene Expression Profiling Services
miRNA Analysis:
Ambion/Affymetrix DiscovArray miRNA Expression Profiling
Agilent Human miRNA Microarray Service
Applied Biosystems TaqMan miRNA Expression Services
DNA Analysis:
Applied Biosystems TaqMan Gene Expression or SNP Detection Services
Data Analysis:
Asuragen’s Informatics Team offers a broad range of analytical services to help you get the most information from your data.
Requirements for Sample Prep Services
Sample Input Requirements for the services listed above are found in the Sample Submission Form and Shipping Instructions.