Bio-bank

The biobank at UMR 8199 is under the responsibility of Philippe FROGUEL. Two technicians are devoted to the biobank: Frederic ALLEGAERT and XXX YYY. The biobank included more than 200,000 biological samples. Its mission is to receive, extract, quantify, maintain and manage the samples.

Thanks to our experience, the laboratory is able to answer to all the services for the preparation and extraction of DNA, RNA from blood, saliva or other tissues and cells.

  • Starlet Hamilton & Autopure LS – QIAGEN
Autopure LS 1Starlet Hamilton

Our robotic platform included 3 DNA extractors, enabling the extraction on a wide range of volume (200μl to 20 ml) of many types of samples, such as fresh or frozen blood samples, saliva samples (via Oragene kit), buffy coat and tissues.

Extraction via the Autopure (Qiagen) is based on a precipitation technique ; this automate enables the extraction of 96 samples per day and automated purification of genomic DNA from large volumes with high throughput, so as to provide high yields, from a wide range of samples (fresh or frozen whole blood, buffy coat, saliva, tissue, …). Incubator / agitator (MAXQ800 Thermo Electron) is also available for rehydration and resuspension of the DNA samples, as well as a pipetting robot that includes a fluorometer, the Biomek NX Span8 (Beckman Coulter).

The extracted DNA samples are then quantitated and standardized (to 200 ng/µl) using a pipetting robot coupled to a fluorometer Beckman Coulter platform, before being incorporated into our database management (LIMS Laboratory Information Management System).

The extraction via Starlet (Hamilton) uses column based purification method – Macherey – Nagel.

These tools allow us to obtain high quality DNA (1.84 +/- 0.02) and stable for their long term storage (biobanking).

 To be kept in a long term periode, the samples are stored in freezers at -80 ° C, and aliquots are placed in a cold room at 4 ° C. These DNAs are available to researchers for their current projects and their external collaborations.

LIMS allows us full traceability and secure, supports through a computerized management of patients, samples and their derivatives.

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  • QIAcube – QIAGEN

The QIAcube (QIAGEN) is a robotic workstation for automated purification of RNA using QIAGEN spin-column kits. Up to 12 samples can be processed per run. All steps in the purification procedure after the disruption and homogenization steps are fully automated. The quality of RNA purified using the QIAcube is comparable to the manual procedure.

In the lab we perform the extraction of:

  • Fiber-rich tissues, such as skeletal muscle, heart, and aorta, tissues difficult to lyse due to the abundance of contractile proteins, connective tissue, and collagen with the RNeasy Fibrous Tissue kit.

We start with samples 10–100 mg to finish with up to 100 µg High-quality RNA eluted in 30–100 µl of RNase-free water water.

  • Fatty tissues, such as brain and adipose tissue with the RNeasy Lipid Tissue Kit.

We start with samples 10–100 mg to finish with up to 100 µg High-quality RNA eluted in 30–100 µl of RNase-free water.

  • Animal or human cells (10 to 1 x 107), animal or human tissues (0.5–30 mg) with the RNeasy Mini kit.

Since the RNeasy procedures enrich for mRNA and other RNA species >200 nucleotides, the total RNA yield does not include 5S rRNA, tRNA, and other low-molecular-weight RNAs, which make up 15-20% of total cellular RNA.

The procedure ensures easy and efficient isolation of high-quality total RNA suitable for any downstream application, including array analysis and real-time RT-PCR.

QIAcube_small_01

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  • Infinite F200 – Tecan
infinite f200

The Infinite® 200 Pro is a spectrophotometer with the ability to perform measurements absorbance and fluorescence. The Infinite 200 PRO can provide a full range of leading detection methods (FRET, BRET) high format flexibility, including 6- to 384-well microplates and to easily and precisely quantify small sample volumes.

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  • LabChip® GX  PerkinElmer

The LabChip GX uses microfluidics capillary electrophoresis for RNA and DNA Quantification.LabChip® GX technology offers unparalleled electrophoresis separation for High Sensitivity DNA/RNA analysis, DNA smear analysis, and RNA and gDNA integrity analysis on one platform.

LabChipGX

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