Biosample Laboratory Services
Nucleic Acid Services
Nucleic acid processing and extraction services are offered for whole blood, biofluids and tissue. We offer flexible DNA extraction capabilities consisting of organic, solid-phase, salting out and bead-based extractions chemistries from virtually any tissue source. We process and extract RNA collected and stored from frozen tissue biopsies, FFPE block/slides, Paxgene RNA tubes, PaxTissue containers, RNAlater and Genotek Oragene RNA collection tubes. We have extensive experience in protocol development and can provide expertise to ensure the right technique is applied for each requisite tissue/sample type.
RUCDR Infinite Biologics uses all methods for DNA extraction including salting out, magnetic beads, solid phase and organic phase. All of these methods can be automated with the exception of the organic method.
- Salting Out
- Magnetic Bead
- Solid Phase
- Qiagen BioRobot Universal - The Qiagen BioRobot Universal is a fully automated nucleic acid extractor capable of isolating DNA from whole blood, buccal swabs and forensic samples. The Universal relies on a unique solid-phase extraction technology that utilizes a filter plate to trap and isolate nucleic acids. DNA is isolated, washed and eluted into a storage buffer. The Universal can process up to 192 samples in 5.5 hours.
- Organic Phase
- Phenol Chloroform - Organic phase extraction remains the gold standard in DNA purification techniques. After lysis, phenol-chloroform is used to separate nucleic acids from proteins and lipids. Precipitation and washing with alcohol results in pure DNA that is resuspended in a storage buffer. Phenol-chloroform DNA extraction is used primarily for old or compromised samples which require special processing.
RNA is extracted using automated solid phase extraction and bead based chemistries, which allows for the isolation/enrichment of micro RNA species (miRNA) in an aliquot separate from the total RNA, within the same workflow. Following extraction, samples are transferred directly into 2D-barcoded tubes and a 2 ul sample for QC is immediately removed by Hamilton STAR automation for analytical and, in some instances, functional QC.
- Magnetic Bead
- Chemagic Star - Manual extraction of RNA is labor intensive and introduces the potential for human error. The Chemagic STAR reduces the risk of error and ensures higher sample quality by: eliminating manual pipetting steps; reducing the risk of cross-contamination; providing faster assay setup; and ensuring consistency of sample processing by removing user variability. The Chemagic STAR utilizes a unique paramagnetic bead technology to purify RNA from PAXgene™ blood. After sample lysis, total RNA is precipitated and bound to the beads. A magnet is used to isolate the beads from solution and wash the RNA before eluting into a special buffer. The Chemagic STAR RNA method isolates total RNA, including smaller types such as miRNA.
- Solid Phase
- Qiagen Universal - The Qiagen BioRobot Universal is a fully automated nucleic acid extractor capable of isolating RNA from PAXgene blood, cultured cell pellets and tissues. The Universal relies on a unique solid-phase extraction technology that utilizes a filter plate to trap and isolate nucleic acids. Total RNA is isolated, washed and eluted into a storage buffer. A secondary protocol is available for the enrichment of miRNA, which normally can be difficult to isolate. The Universal can process up to 192 samples in 5.5 hours.
- Organic Phase
- Phenol Chloroform - Organic phase extraction results in high purity RNA. After lysis, phenol-chloroform is used to separate nucleic acids from proteins and lipids. Precipitation and washing with alcohol results in pure RNA that is resuspended in a storage buffer. Phenol-chloroform RNA extraction is used primarily for old or compromised samples which require special processing.
Nucleic Acid Quality Control
After extraction, all nucleic acid samples are subject to comprehensive analytical and functional quality control procedures. Analytical quality control consists of concentration, purity and volume measurements utilizing cuvetteless spectroscopy that measures contaminant profiles allowing for precision measurement of nucleic acid. Functional quality control consists of the RUID™ quality panel for DNA. Our quality control provision is state of the art and in place to ensure sample quality and integrity for downstream analysis.
- Analytical Quality Control
- Trinean DropSense96 - Concentration is measured using the Trinean DropSense96. The DropSense96 is a multichannel spectrophotometer that uses UV/VIS spectral analysis to quantify microliter droplets of DNA or RNA. Data generated from the DropSense 96 is highly robust and measure contaminates, as well as double stranded DNA as low as 5 ng/ul.
- Functional Quality Control
- RUIDGX™ - RNA is quality controlled using the RUIDGx™ quality panel. This approach analyzes tissue-specific transcripts that interrogate the entire length of selected transcripts as well as a variety of expression levels. This rapid analysis provides direct assessment of cDNA fidelity and provides a score for each sample which correlates directly with microarray and RNAseq performance for any sample.
Nucleic Acid Amplification
RUCDR offers multiple solutions for the robust amplification of precious nucleic acid samples, as well as restoration of degraded or compromised material. Nanogram quantities of starting material are converted into microgram quantities for storage or downstream applications. Whole-genome and whole-transcriptome amplification chemistries enable highly uniform and unbiased enrichment, preserving relative abundance and allelic balance for downstream genetic analyses. Amplified template is suitable for qPCR, microarray and Next-Generation Sequencing applications.