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
Automation
Where possible, Asuragen offers automated solutions for RNA and DNA sample preparation,
using the KingFisher® 96 robot from Themo Scientific. The King Fisher®
is designed to automate the time-consuming preparation process of nucleic acids.
Implementation has increased batch size of PAXgene® blood processing
4-fold, with yields and quality equivalent to the manual protocol.
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
for mRNA analysis are routinely qualified by spectrophotometry with the NanoDrop spectrophotometer and the Agilent 2100 Bioanalyzer. All samples prepared by Asuragen
Services for microRNA analysis are routinely qualified by spectrophotometry with
the NanoDrop spectrophotometer
and qRT-PCR assessment of discrete miRNAs is used as a surrogate measure of overall
miRNA abundance. 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 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.
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.
Requirements for Sample Prep Services
Sample Input Requirements for the services listed above are found in the Sample Submission
Form and
Shipping Instructions.