Policy

General Information

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.

Protein Expression

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.

 

 

Robotics

The Molecular Biology Service offers different automated liquid handling systems to the scientists of IMP, IMBA and GMI.

After an introduction on the systems, users are allowed to use the robots to pipette their experiments like:

  • PCR set up
  • Real Time PCR
  • Nucleic Acid Purification
  • High Through Put In Situ Hybridizations

 

We together with new users program new automated workflows.

As a routine service we offer automated DNA extraction (Plasmid and genomic DNA).

 

 

 

Sanger Sequencing

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.

 

Sample concentration:

-          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:
\\storage.imp.ac.at\groups\MolBioService\CollaborationData\SEQUENCINGRESULTS-new (PC)

or smb://storage.imp.ac.at/groups/MolBioService/CollaborationData/SEQUENCING RESULTS-new (MAC)

SAMPLE DEADLINE

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.

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.

Mycoplasma Tests

The procedure is guided by an SOP. Routine mycoplasma testing service for tissue culture cells is established since September 2012, based on the method described in the Australian Patent “Assay for detecting mycoplasma by measuring acetate kinase or carbamate kinase activity”.

Deliver 250µl of your samples on Thursday at 11am in the Lab 2-012.

Microarray

The Molecular Bilogy Service uses Gene Expression Microarrays and CGH Microarrays from Agilent. 

Initial Meeting: Before we start a microarray experiment it is necessary to meet and discuss following items:Minimal Information about a Microarray Experiment (MIAME), experimental design and addressed questions. Download: MIAME 

The actual Experiment is done by Harald. We start the experiment after we confirmed the quality using the Bionanalyzer from Agilent. We introduced several quality control checkpoints in our experimental work flow. This enables us to stop an experiment before we hybridize them onto the arrays.

  • Analysis and Closing Meeting:

The scientist is given an Access database containing all result tables of interest. These primary results can be further analyzed e.g. on the workstation with Spotfire Decision Site data analysis.

Media Kitchen

The Media Kitchen 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 or come to our facility to talk about your personal special requirements. You can download the list with available products: Stock List