The Molecular Biology Service is an open lab where members of the institutes IMP, IMBA and GMI can use most of the lab equipment after a proper introduction. After the introduction you can use the devices at any time. Note, some of the devices are bookable through the Booking System (http://grm3.imp.univie.ac.at/). Users are responsible for leaving the workstations clean and in working order. In case we observe constant misuse or not properly cleaned devices, we will make restrictions on use of the equipment. In case you encounter any problems please report them to us.
The use of recombinant proteins in daily scientific research has increased greatly in the last years. The Protein Expression Facility is established to provide support in all aspects of protein expression and purification.
Our main goals are:
Establishing and maintaining collections of expression vectors and host strains.
Providing low-cost recombinant proteins for researchers (such as proteases, polymerases and growth factors).
Elaborating and testing new protocols for protein expression and purification as well as keeping track on the newest developments in this area.
Offering technical assistance and training to researchers for successful production of their recombinant proteins.
The Molecular Biology Service offers different automated liquid handling systems to the scientists of IMP, IMBA and GMI.
Agilent Bravo Automated Liquid Handling Platform
After an introduction on the Bravo (Agilent) systems, users are able to use the robots to pipette their experiments like:
· - Rapid, high-precising PCR setup
· - different bead applications in 96 well format
· - 96well liquid handling for DNA purification
Tecan clone handling robot
The Evo75 liquid handling platform (Tecan) in combination with a Pickolo clone picking module (Scirobotics) will expand and streamline our service of DNA isolation with robotic assistance. The robot is equipped with a gripper arm for plate handling and 2 steel tips for liquid handling that can be decontaminated from inside and outside thereby replacing the use of plastic tips. Moreover the robot has the colony picking module including a light table, CCD camera and the Pickolo software for colony identification. The carrier setup can hold 24 9cm petri dishes, 6 MTP plates on main platform and additional 10 MTPs in the hotels.
Main competences of the current setup are
- Colony picking, e.g. into 96 deep well plates for plasmid preparations
- Colony picking for colony PCR
- Cherry picking
- Colony counting
King Fisher Duo (Prime)
The KingFisher instrument has a comb with 12 magnetic rods to transfer magnetic beads into different solutions using a sterile plastic comb. The device can be used for various kinds of applications where magnetic beads are subjected to different solutions, e.g. RNA isolation DNA isolation, IPs etc. On deep-well row can be heated, the elution strip can be heated and cooled. The KingFisher will help you to reduce hands on time and achieve high reproducibility. It can be used as stand-alone device or connected to a PC when editing of protocols is wanted. The intuitive software allows easy customization of protocols.
· 2 magnet combs for 6 or 12 samples
· 2 positions for deep well plates and elution strips for up to 24 samples
· 30- 5000 ul volume
· High reproducibility
· UV-lamp for decontamination (Duo Prime)
· Very easy to use
· Intuitive software to customize protocols
· Heating row A: up to 75°C
· Elution strip: 4-75°C
· Can be operated in cold room
Routine service offered
As a routine service we offer automated DNA extraction (Plasmid and genomic DNA).
The Sanger Sequencing Facility provides automated sequencing using capillary based AB3730 and AB3730XL DNA analyzers. These instruments offer high throughput and sensitivity with a capability to handle 800 to 1600 samples per day. These analyzers allow us to have turnaround time of 1 to 2 days. Samples of single stranded, double stranded DNA, and PCR products, can be sequenced with >800 bps and a success rate of over 90%.
Sample Submission Guidelines
Bring your samples and primers to room 2-012 at the 2nd floor in IMP building and put them in the small glass door refrigerator. Put samples into racks labeled as “SAMPLES” and primers into racks labeled as “PRIMERS”. Some primers are commonly available in the Facility, please see LIST OF PRIMERS. Use the Sanger Sequencing Sample Submission software to give us all information about your samples.
- 150 – 200 ng for plasmid DNA per reaction
- 20 – 50 ng for PCR products per reaction
- 500 – 1000 ng for cosmid, BACs and YACs per reaction
* Individual samples prepare in to 1.5 ml tube or 8-well stripes
* If you have 16 or more samples (it means, e. g. 10 samples with 2 primers), please prepare them in a plate (see PLATE PREPARING).
* In a case of samples containing secondary structures, GC rich regions or shRNAs please ask for “secondary structure protocol”
* Results from sequencing – you get email with the links or you can download them from:
or smb://storage.imp.ac.at/groups/MolBioService/CollaborationData/SEQUENCING RESULTS-new (MAC)
Samples must be brought to the room 2-012 IMP by 15 pm to be run that day.
“Speed Congenics” Service
Genetically modified mouse are mainstream tools for medical research. In many cases, the background strain used to create the genetic modification is inappropriate for phenotypic analysis of the mutation. In such cases it is useful to develop a congenic strain of the transgenic mouse line in which the mutation is introduced into a more suitable genetic background. By targeting selection of the mouse with the highest percentage of the receiver’s genome at each backcrossing generation, the congenic strain can be developed within 4-5 backcross generation instead of 10 generation by using a traditional strategy.
Our facility offers a simple sequence length polymorphisms (SSLP)-based screening of the mouse genetic background. The screening is achieved by using specific sets of more than 90 markers evenly dispersed in a distance of 20 cM (≈ 20 - 40 Mbases) over the 19 autosomal mouse chromosomes.
Speed congenic service is offered now for commonly used mouse strains combinations: C57BL/6J versus 129, C57BL/6J versus BALB/c, and C57BL/6J versus FVB/n, and C57BL/6J versus CBA. We are also able to distinguish between C57BL/6J (Jackson) and C57BL/6N (NIH), necessary because of different phenotypes. According to request can be customized for other mouse strain pairs in 5-8 weeks. Please download request sheet.
Authentication of Human Cells
Unambiguous cell authentication is very important for credibility and reproducibility of the scientific results and is ask for review of papers submitted for publication or for review of grants submitted for funding.
STR (short tandem repeat) profiling for authentication of human cell lines, stem cells and tissues is done by capillary based analysis of 16 STR markers as is recommended by Standard ASN-0002 (ATCC). The results for cell lines are compared with ATCC or DSMZ databases.
How to get your samples tested:
Contact Martina Rath per email (martina.rath(at)imp.ac.at) and then bring around 10 ul (concentration 50 ng/ul) of isolated DNA to our lab (IMP 2-012).
Fragment Analyzer and ddPCR System
Robert is the person to ask for help and training on the Fragment Analyzer and ddPCR System.
The Fragment Analyzer™ is a powerful instrument to cover the full spectrum of DNA, gDNA and RNA quality control by parallel capillary electrophoresis with fluorescence detection. Applications range from standard nucleic acid separations and quantification across the widest separation range to high-resolution analysis. In our Lab we provide 12 and 96 capillary instruments.
The QX200 Droplet Digital PCR (ddPCR™) System provides absolute quantification of target DNA or RNA molecules for EvaGreen or probe-based digital PCR applications.
A PCR reaction is set up comparable to real time experiments and divided into several thousand individual subreaction by generating droplets where only a fraction of droplets contain target sequence. After thermal cycling (endpoint PCR) each droplet of a reaction is analysed in a digital way (signal, no signal) directly leading to an absolute quantification.
The MBS provides routine service for examine mycoplasma contamination in cell cultures. This service is established since September 2012. In the first 5 years, the test was performed according to the method described in the Australian Patent “Assay for detecting mycoplasma by measuring acetate kinase or carbamate kinase activity”. We have switched from enzymatic system to real-time PCR based method since June 2017, as described in “A Single-Tube Real-Time PCR Assay for Mycoplasma Detection as a Routine Quality Control of Cell Therapeutics”, by Janetzko K, et al. This procedure enables rapid and specific detection of mycoplasma contamination by using mix of primers, that are specific to conserved region in 16S rDNA sequence. We have developed above assay and increased the number of Mollicutes species, which can be determined up to 220.
PCR is performed on cell culture samples provided by users and results are obtained within few hours on the submission day.
How to get your samples tested:
Bring your samples as usual on Thursday latest at 11am to our laboratory. In case Thursday is a public holiday, we do the test on Wednesday.
The Media Lab is the core facility of media preparation. Its aim is to produce high quality media. It provides a variety of common stock products and is always eager to serve you with personal tailored solutions. Feel free to order high amounts of standardized media with the online ordering sytem or come to our facility to talk about your personal special requirements. Ordering System for Media and Buffers.