Sample Preparation and Isolation of RNA or DNA
Blood and Other Biofluids
As early as 2003, Asuragen scientists (then at Ambion) began developing methods to isolate RNA from cell-containing and cell-free biofluids. This work is now reflected in several successful Ambion products, but Asuragen scientists have worked to further enhance these methods, and Asuragen Services now has experience with these protocols in clinical settings.
Biofluids, such as blood, serum, plasma, urine, saliva, cerebrospinal fluid (CSF), and others can hold the key to unlocking valuable information in the discovery and development of biomarkers. These biomarkers could, in turn, improve the prediction of clinical trial failures and decrease the costs associated with failed drugs.
Asuragen has developed and qualified isolation procedures to extract nucleic acids (RNA and DNA) from these unique samples, and can assist in review of collection methods, collection site training, and even sample collection kit configuration.
Learn more about how we qualify our methods and staff.
Description of Services
Deliverables
Quality Control
Turn-around Time
Typical Yields
Additional Services Available
Catalog Numbers for Blood/Biofluid Sample Prep Services
Description of Services
Sample prep: Blood
Features of blood sample prep services:
- Total RNA isolation from whole blood using PAXgene®, including recovery of the miRNA fraction using proprietary isolation procedures
- Optional globin message reduction
- Methods may utilize exogenous DNA or RNA spiked-in controls
Workflow for blood sample prep:
- Total RNA isolation from whole blood using PAXgene, including recovery of the miRNA fraction using proprietary isolation procedures
- Optional globin message reduction
- Methods may utilize exogenous DNA or RNA spiked-in controls
Sample prep: Other biofluids
Workflow for other biofluids sample prep:
- proprietary procedures for isolation of RNA and DNA from other biofluids unique to sample type
- QC by NanoDrop spectrophotometer
- QC by Bioanalyzer is not standard for biofluids
- Optional QC measures can be used to indicate suitability for mRNA or miRNA applications
- All RNA isolation methods optimized to recover miRNA
- Asuragen-isolated RNA or DNA is suitable for numerous downstream applications, including expression profiling and genotyping studies
Globin reduction service for whole blood
Globin mRNA from red blood cells makes up the majority of RNA in whole blood total RNA samples. If RNA samples from whole blood are to be used for array-based gene expression profiling, the presence of globin mRNAs can lead to diminished array sensitivity, increased background noise, and an overall reduction in the number of genes called “present” [1]. To address this, Asuragen can assist with special protocols at collection for highly efficient removal of red blood cells or specific removal of the abundant globin transcripts after the total RNA has been isolated.
Deliverables
Regardless of downstream application, detailed QC reports including nucleic acid concentration, RNA integrity (RIN) and electropherograms where applicable, and purity are provided on each sample. Purified nucleic acid samples are shipped back to you on dry ice. Asuragen ships samples Monday through Wednesday and will notify you upon shipment.
Isolation services are often utilized with Asuragen’s molecular profiling services. Surplus RNA or DNA will be discarded unless otherwise requested. Additional shipping charges may apply for the return of residual blocks, RNA, or DNA.
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. 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.
High Quality RNA
RNA was isolated from colon tumor tissue by Asuragen and run on the Agilent 2100 BioAnalyzer to show integrity (red line). The same RNA was incubated overnight at 37°C to show stability (blue line). This data shows that there was no change in RNA quality and integrity after extended incubation.
These 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 commitment to the best possible data quality, or read about our contributions to in-process controls and standardization.
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
Typical Yields
Over the span of a year, Asuragen processes thousands of PAXgene blood samples for clients in the pharmaceutical industry. Our PAXgene-based protocol optimized for microRNA recovery yields high-quality RNA suitable for numerous downstream applications. Asuragen also has experience isolating nucleic acids from a variety of biofluids and has developed suitable methods for both mRNA and microRNA applications.
Sample type |
RNA yield/mL |
blood |
<1 to >10 mg |
plasma/serum |
5-50 ng |
saliva |
100 ng |
urine |
3-20 ng |
Yield, integrity, and purity of RNA are influenced by a variety of endogenous and exogenous factors, including:
- Sample composition
- Mass or volume
- Disease status
- Chemical treatment
- Preservation upon collection
- Sample handling
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.
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.
.
Catalog Numbers for Blood/Biofluids Sample Prep Services
| CA0045 | Isolation, PAXgene Blood, Total RNA, with miRNA and in-process spike |
| CA0022 | Globin Reduction Blood Total RNA (mass sensitive: 10 mg per sample max) |
| CA0025 | Isolation, DNA, Custom |
| CA0010 | QC, NanoDrop and Bioanalyzer, Total RNA, for mRNA Applications (applies to blood) |
| CA0032 | QC, NanoDrop and TaqMan, Total RNA, for microRNA Applications |
| CA0037 | QC, NanoDrop and TaqMan, Total RNA, for mRNA PCR (applies to biofluids, FFPE) |
| CA0036 | Isolation, Total RNA, Custom (applies to other biofluids) |
Requirements for Blood / Biofluids Sample Prep and Related Services
Sample Input Requirements for the services listed above are found in the Sample Submission Form and Shipping Instructions
References
1. Madabusi, LV et al., RNA Extraction for Arrays. Methods Enzymol, 2006. 411: p. 1-14.