These machines are used as a self-service and you will require prior training by one of the members of your team. For new teams arriving at the Institute, this training will be offered by Thierry Grange or a member of his team.
The machines are usually very busy. You must therefore reserve in advance using the notebook available by each machines. Unless you need to use a lot of cycles (>50), it is mandatory to reserve - at the most - a 90 minute session and to respect it. Your samples should normally be prepared before you start your session.
Roche Light Cycler 1.5
The Light Cycler 1.5 allows the reliable, sensitive and specific quantification of nucleic acid sequences, the characterisation of PCR products by melting curve analysis and genotyping of SNP in less than one hour. It is possible to analyse up to 32 20-µl glass capillaries simultaneously. Sample excitation can be achieved with a maximum emission at 470 nm, and fluorescence can be measured simultaneously at 530, 640, and 705 nm. Measurements can be done with SyBr Green I, specific probes (Taqman, Scorpion, Beacon, etc…) labelled with FAM, and hybridization probes labelled with the couples FAM-LightCycler Red 640 or 705. Multiplexing is not possible except in the hybridization probe format.
For a more detailed description, see http://www.roche-applied-science.com/sis/rtpcr/lightcycler/index.jsp?id=lct_010502.
Roche Light Cycler 480
The Lightcycler 480 is a fully integrated multiwell-plate based real-time PCR platform for highly accurate qualitative and quantitative detection of nucleic acids. The available device is equipped with a 384-well block. The high-performance silver thermal block cycler built into the LightCycler® 480 System enables completion of a PCR run within one hour (384-well block), simultaneously enabling maximized homogeneity during heating and cooling over a broad temperature range for many targets and applications without sacrificing well-to-well homogeneity.
It offers a large choice of excitation and detection wavelength for versatile applications including multiplexing and high resolution melting curve analyses.
|Assay/Detection Format||Dyes||Excitation (nm)||Detection (nm)|
|SYBR Green l||SYBR Green I||465||510|
|Hydrolysis Probe / UPL Probe Monocolor||FAM||465||510|
|Hydrolysis Probes / UPL Probes|
VIC / Hex / Yellow 555
VIC / Hex
Cy 5 / Cy 5.5
|LightCycler® Cyan 500|
LightCycler® Red 610
Cy 5 / Cy 5.5
LightCycler® Red 640
LightCycler® Red 610
LightCycler® Red 640
Cy 5 / Cy 5.5
For a more detailed description, see https://www.roche-applied-science.com/sis/rtpcr/htc/index.jsp.
This 96-well QPCR is slower than the Roche 480 but offers access to the 96 well format. It is equipped with 5 filters allowing the independent or simultaneous use of FAM/SYBR Green 1, HEX/JOE/VIC, ROX/Texas Red, Cy5, and Cy3.
There is also a Stratagene MX3000 P device available that is currently hosted in Reiner Veitia's group allowing the use of 4 filters.
For a more detailed description, see http://www.genomics.agilent.com/CollectionSubpage.aspxPageType=Product&Sub...
Pipetting and preparation Robots
Eppendorf epMotion 5070
The epMotion 5070 allows accurate and reproducible automated pipetting. With 4 deck positions, it can hold tips, reagent reservoirs, single tubes, tube racks, micro-plates, or PCR plates with up to 384 wells. It allows routine application such as serial dilutions, reagent distribution, sample transfer from tubes to plates, and sample normalization. It is particularly useful for PCR setup using the 384 and 96-well formats. It is currently equipped with a either a single or 8 channel 50 µl pipettes.
For a more detailed description, see http://www.eppendorf.com/int///index.php?pb=d74b4ccde117f841&page=1&action=epmotion&contentid=3&page=2.
Freedom Evo 100
This is a versatile robot for high throughput sample preparation equipped with a 4 channel liquid handling arm suitable for disposable tips, a gripper to move objects, a thermoblock shaker and an automated solution for vacuum solid phase extraction that can be assembled by the robotic manipulator arm for fully automated operations that is suited for DNA /RNA extraction and other purifications.
For a more detailed description, see http://www.tecan.com/platform/apps/product/index.asp?MenuID=2694&ID=5270&Menu=1&Item=21.1.8.
Microarray hybridization and scanning
Hybridization station Tecan HS400 Pro
The HS 400 Pro microarray hybridization stations minimize handling of microarray slides by implementing all the steps of microarray processing, from pre-hybridization and blocking to washing and drying, inside the slide-processing module., The microarray hybridization stations are fully programmable with accurate time and temperature control, and tunable frequency of sample agitation to match the requirements of applications ranging from hybridization of high-density DNA arrays to processing of protein arrays. The HS 400 Pro can handle up to 4 slides.
For a more detailed description, see http://www.tecan.com/platform/apps/product/index.asp?MenuID=1791&ID=1840&Menu=1&Item=21.10.1.
Nimblegen Hybridization system 4
This hybridization station allows processing of Nimblegen high-density microarrays.
For a more detailed description, see http://www.nimblegen.com/products/instruments/hybridization/4-slide/index.html.
High density microarray scanner Tecan Power Scanner
The PowerScanner allows scanning of high-density DNA microarrays and protein microarrays with maximum resolution of 2 micron/pixel. It is a dual channel microarray scanner fitted with powerful green and red lasers and emission filters for excitation and detection of commonly used fluorophores. It is able to identify and focus automatically on the arrayed surface of glass slides, including nitrocellulose-coated and reflective surfaces. It includes an internal autoloader with capacity for up to 48 slides, and air filters to minimize ozone inactivation of fluorophores
For a more detailed description, see http://www.tecan.com/platform/apps/product/index.asp?MenuID=3241&ID=6444&Menu=1&Item=21.10.5.
96-well plate readers and other analyses equipment
Absorbance reader Dynex MRX
The MRX Microplate Reader is a microprocessor-controlled photometer that t is designed to measure the optical density (OD) of fluid samples in 96-well microplates in order to determine the level of chemical reactions. Spectral Range 400 to 850nm. Installed 3 standard onboard filters 405, 450, 492nm or user's choice of filters. Single/dual or multiple wavelength readings; dual wavelength <6 seconds, single wavelength reads <4 seconds. Endpoint, kinetic plate reads to 1000 per plate, plate shaking, calculation range. Dynamic range -0.100 to 4.000 OD.
For a more detailed description, see http://www.cox-scientific.com/Images/Product%20Images/Reconditioned/Manuals/DYNEX-MRX_II_Reader_User_Manual.pdf.
Microplate Luminometer Berthold Centro LB 960
The Centro LB 960 is a versatile, robust and sensitive microplate luminometer that supports all basic luminescence technologies including Flash-type Luminescence, Glow-type Luminescence, Cellular Luminescence, Bio-Luminescence and Chemi-Luminescence. It is equipped with 2 injectors.
For a more detailed description, see https://www.berthold.com/en/bio/product/centro-lb-960-microplate-luminometer.
Microplate spectrophotometer and fluorimeter Tecan Infinite 200 Pro
The Infinite 200 PRO uses advanced optics and high-performance detectors, optimized for the requirements of fluorescence and absorbance reading. The Infinite 200 PRO offers a wide range of detection modes, including: Fluorescence intensity (top and bottom reading) (UV – NIR & wavelength scanning), Fluorescence resonance energy transfer (FRET), Time resolved fluorescence energy transfer (TR-FRET) like HTRF, Time resolved fluorescence (TRF), Fluorescence polarization (FP), Absorbance (UV – NIR & wavelength scanning), Temperature control and Small sample measurement in NanoQuant Plate.
For a more detailed description, see http://www.tecan.com/platform/apps/product/index.asp?MenuID=1812&ID=1916&Menu=1&Item=126.96.36.199.
Microspectrophotometer Nanodrop ND1000
The Nanodrop ND1000 allows acquisition of absorbance spectra between 220 and 750 nm for 1µl samples that are held in place using surface tension.
For a more detailed description, see http://www.nanodrop.com/library/nd-1000-v3.7-users-manual-8.5x11.pdf.
Lab on a chip electrophoresis: Agilent 2100 Bioanalyzer
The Agilent 2100 Bioanalyzer is a microfluidics-based platform for sizing, quantification and quality control of DNA, RNA, proteins and cells. Results are delivered within 30-40 minutes in automated, high quality digital data. Miniaturization of analytical instrumentation has many advantages over conventional techniques. These advantages include improved data precision and reproducibility, short analysis times, minimal sample consumption, improved automation and integration of complex workflows.
Area of application: RNA quality check with RIN (RNA Integrity Number), RNA analysis of total RNA, mRNA and Small RNA; DNA size and quantity, high resolution separation and quantification of DNA, highly sensitive DNA sample QC for next generation sequencing experiments; SDS-PAGE replacement for protein analysis to determine the quantity and purity of proteins from Coomassie down to silver stain sensitivity.
For a more detailed description, see http://www.genomics.agilent.com/CollectionOverview.aspx?PageType=Platform&SubPageType=PlatformOverview&PageID=275.
Lab on a chip electrophoresis for DNA sizing and purification: Caliper Labchip XT
The LabChip XT fractionation system performs fast, automated nucleic acid fractionation accurately and reproducibly. The resulting sample is tightly sized and is delivered in a sequencing compatible buffer. The XT and provides efficient sizing that is difficult to obtain using manual methods. Data is displayed digitally and non-fractionated sample can be recollected and used at another time. The XT has advanced extraction modes and is ideal for users with more complex applications with data analysis and sample tracking needs. It is particularly adapted for sizing of Next Generation sequencing libraries.
For a more detailed description, see http://www.caliperls.com/products/labchip-systems/labchip-xt.htm.
Next generation sequencing
Ion Torrent Personal Genome Machine
Powered by Ion Torrent™ semiconductor chip technology, the Ion Personal Genome Machine™ (PGM™) Sequencer delivers the fastest run times, at the most affordable price, of any next-generation sequencer. The Ion PGM™ Sequencer is ideal for sequencing small genomes, sets of genes, or performing gene expression profiling or ChIP-Seq. Its particular features are its speed, sequencing run times last about 2 hours, the possibility to perform sequencing at low costs using inexpensive reagents and its scalability (1000x) using semiconductor chips of various capacities. With the Ion 314 Chip: up to 100,000 reads or up to 20 Mb, with the Ion 316 Chip: up to 1 million reads or 200 Mb, Ion 318 Chip: up to 5 million reads or 1 Gb.
For a more detailed description, see http://products.invitrogen.com/ivgn/product/4462917.
Illumina MiSeq personal Sequencer: Available soon!
MiSeq uses TruSeq, Illumina's reversible terminator-based sequencing by synthesis chemistry. The Miseq allows to perform standard experiments such as amplicon sequencing, clone checking and small genome sequencing with enhanced performance, more quickly, and cost-effectively compared to capillary electrophoresis (CE) sequencing. Suitable for next-generation sequencing applications such as highly multiplexed PCR amplicon sequencing, targeted resequencing, ChIP-Seq, small RNA sequencing, and more. The format is also particularly adapted for rapid verification and optimization of libraries that can be subsequently used on high performance sequencers like HiSeq. Up to 15-17 Millions reads can be sequenced at a time.
Approximate Run Duration and Output
Cluster Generation and Sequencing
|Read Length (bp)||Total Time*||Output|
|1 × 36||~4 hrs||540-610 Mb|
|2 × 25||~5.5 hrs||750-850 Mb|
|2 × 100||~16.5 hrs||3.0-3.4 Gb|
|2 × 150||~24 hrs||4.5-5.1 Gb|
|2 × 250||~39 hrs||7.5-8.5 Gb|
For a more detailed description, see http://www.illumina.com/systems/miseq.ilmn.