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

Features of blood sample prep services:

  • Nucleic acid isolation from whole blood collected in PAXgene® tubes
  • Recovery of the miRNA fraction using proprietary isolation procedures, if applicable
  • Optional globin message reduction

Automation

Workflow for blood and other biofluids:

  • Custom isolation service (see above for features of the blood sample prep service)
  • Proprietary procedures for isolation of RNA and DNA from blood and other biofluids unique to sample type
  • QC by NanoDrop spectrophotometer
  • Optional QC measures can be used to indicate suitability for mRNA, miRNA, or DNA 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 gene expression services, miRNA profiling services, or genotyping 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.

For RNA

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

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.

For DNA

All DNA samples prepared by Asuragen Services are routinely qualified by spectrophotometry with the NanoDrop spectrophotometer and gel electrophoresis. Due to the degraded nature of DNA from FFPE tissues, special quality control measurements are used.

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 one to two weeks. 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 ug
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
Applied Biosystems TaqMan Gene Expression Services

miRNA Analysis:

Ambion/Affymetrix DiscovArray miRNA Expression Profiling
Agilent miRNA Microarray Service
Applied Biosystems TaqMan miRNA Expression Services

DNA Analysis:

Agilent aCGH
Affymetrix SNP 6.0 - coming soon

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
CA0022 Globin Reduction Blood Total RNA (mass sensitive: 10 ug per sample max)
CA0025 Isolation, DNA, Custom
CA0057 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.