Hope Center DNA/RNA Purification Core

The goal of the DNA/RNA Purification Core is to facilitate genetic and multiomic analyses by providing services for sample collection, processing, storing and data generation.

Sample Collection Kits and Biobanking: Starting January 3^rd, 2023, the Hope Center DNA/RNA Purification Core also provides support for sample collection and banking. This encompasses designing and a continuous supply of collection kits specific for blood, buffy, plasma, PAXgene and PBMC processing for ongoing or future projects. The core can also coordinate shipping and returning of collection kits, along with sample processing, and storage for the collected samples. All kit collection and storage tubes are 2D barcoded for ease of tracking samples.

DNA and RNA extraction: The DNA/RNA purification core has instruments that allow the extraction of nucleic acids from a larger variety of tissues, at high throughput, and at a competitive price.

We offer DNA and RNA insolation from whole blood (large or small volume), buffy, buffy coat, saliva, buccal swab, blood cards, plasma, cultured cells, or tissue.

We also offer DNA and RNA QC, low and medium throughput SNP genotyping (1-30 SNPs), APOE genotyping, Sanger sequencing,  DNA/RNA normalization, and DNA/RNA plating for transfer to GTAC@MGI (or another service provider) for genotyping or sequencing.  Recently added services include controlled temperature plasma and CSF aliquoting for OMIC data generation, extended BioBank sample storage, and PBMC isolation.

We now offer PBMC isolation and concentration measurements from whole peripheral blood collected within the last 24 hours utilizing in-vitro density gradient centrifugation along with SepMate technology and the Countess III.

For a complete list of services and current prices please check the Current Fee Schedule

Guidelines for Submitting Samples

Following the suggested collection, preparation and transport instructions below supports the best possible test results. Plastic collection tubes are required for all samples.

Please contact Kristy Bergmann (k.bergmann@wustl.edu) or Pat Kohfeld (pkohlfeld@wustl.edu) and fill out the Sample submission form before dropping off samples.

General notes

For best results, freshly drawn blood should be stored at 4°C for less than 4 days, or should be frozen at   -80°C on the day of collection if possible.  Samples should be submitted to the core within 1 – 4 days after collection for optimal results.  Samples at can be also storage at -20°C but for no more than 3-6 months.

All sample tubes should be labeled with a unique sample identifier, the PI name, and the date collected.

DNA/RNA will be returned in 2D barcodes tubes, unless a different tube is requested in advance.

NanoDrop information will be provided for all samples extracted by the Hope Center DNA/RNA core.

Whole Blood and buffy

Blood samples should be collected in PLASTIC TUBES, preferably EDTA (e.g. BD Vacutainer #366643 purple cap tubes). Collection in ACD (citrate, yellow cap tubes) is also acceptable. Please DO NOT USE GREEN CAP (Heparin) Tubes. Other plastic blood collection tubes are acceptable provided they meet the guidelines above.

Tissue and cell pellets

Tissue samples and cell pellets should be snap frozen on dry ice and stored at -80°C.  Samples should never be stored at -20°C as they quickly degrade.

Samples stored for more than 1 year or at less than -80°C may result in lower yield and quality of DNA.  Multiple freeze/thaws will also affect DNA yield and quality.

Resources

Autogen FlexSTAR+ Automated Workstation

Large volume DNA samples (1 – 10 ml) are extracted on the Autogen FlexSTAR+ using proven Flexigene chemistry, a salt precipitation method that removes protein contaminants and enzyme inhibitors. The FlexSTAR+ platform is capable of high-throughput purification from large volume samples (1 – 10 ml) in ~ 4 hours (30 samples for 1-5 mL or 15 samples for 6-10 mL.)

Maxwell RSC 48 Automated Workstation

DNA or RNA are extracted from small volume samples (generally < 500mL) using magnetic bead-based purification chemistry. The Maxwell RSC 48 can process a large variety of input sources (blood, buffy, saliva, blood cards, tissue, buccal swab, plasma, CSF, cultured cells, and others).

Liquid handling workstations

The Biomek FXP liquid handling automated workstation enables high-precision pipetting, eliminating manual pipetting errors and ensuring consistent, reproducible results. The Biomek FXP includes a multichannel (96 head) and a Span-8 head. This increased efficiency and flexibility all allow high-throughput protocols.

Agilent TapeStation 4200 System (DNA/RNA Quality)

The TapeStation 4200 system is an established automated electrophoresis tool for DNA and RNA sample quality control.  DNA and RNA quality and quantity are assessed with the TapeStation 4200, which is able to run up to 96 samples in one run and provides both concentration and quality metrics (DIN or RIN). Fully automated sample processing enables the unattended analysis of size, concentration and integrity. The TapeStation system provides a complete solution for true end-to-end sample quality control within any next-generation sequencing (NGS) or Biobank workflow.

ABI3500 Automated DNA Sequencer

The 3500 Genetic Analyzer is an 8-capillary sequencing instrument specifically designed to support the demanding performance needs of validated and process controlled environments, while retaining the unsurpassed application versatility that life science researchers expect. The 3500 Series Data Collection Software supports sequencing and fragment analysis, and integrates seamlessly with downstream software for secondary analysis of genetic data.

Real-time PCR instruments

The Life Technology StepOnePlus Real Time PCR machine (96-well plate) and a QuantStudio 12K Real Time PCR machine (384-well plate) are used for qPCR, allele specific gene expression, and SNP genotyping.

Countess III (Cell Counting System)

Thermo Fisher Scientific Countess 3 Automated Cell Counter provides accurate and precise brightfield and florescence cell counts. Its automated features like rapid capture, autosave, and built-in calculators add convenience and time savings for counting peripheral blood mononuclear cells (PBMCs).

AFFILIATIONS
Hope Center for Neurological Disorders (HCND)