The Molecular Biology and Protein Expression Automated Platform, RoBioMol, is hosted by the Pneumococcus Group of the Institut de Biologie Structurale (IBS) and offers high throughput molecular biology processes using automates.
Current services include: - gene cloning, - site-directed mutagenesis, - expression and purification test of proteins expressed in E. coli - automated plasmid preparation. These services are available for a minimal number of cloning / expression tests of 24 samples. Data are processed and managed via a Laboratory Information Management System (RoBioLIMS) specifically developed for the platform by the CEA (GIPSE).
How to make a request?
Contact the RoBioMol team (firstname.lastname@example.org) to submit your request.
For gene cloning and site-directed mutagenesis, the gene to be mutated or cloned should be provided either cloned into a plasmid (10 ng minimum/cloning) or as a chromosomal template (ie: bacterial DNA, 100 ng/cloning). The
quality of the DNA template will be controlled by the RoBioMol team
(concentration and 260nm/280nm ratio determination, analysis by agarose
For plasmid minipreparation, bacterial colonies containing the plasmids to be prepared should be provided in petri dishes at RoBioMol place.
For detergent screening for membrane protein solubilization and detergent exchange, membrane preparation, which has to be previously analysed to ascertain the presence of over-expressed His-tagged protein, has to be put down at RoBioMol place.
Samples must have security level L1 and conform to the corresponding GMO statement (GMO agreement number is required).
Should the quantity or quality of the matrix departs from the RoBioMol standards, the service will be delayed and the requesting scientist will be contacted.
Template will be renamed for internal use, stored frozen for plasmid or genomic DNA and membrane preparation, or between 0°C and 10°C for bacterial petri dishes. Template will be discarded at the end of the work unless specified otherwise on the order form.
Gene cloning and site directed mutagenesis deliverables:
the plasmid DNA containing the cloned or mutated gene is delivered in
solution (a few nanogram, enough to perform a bacterial transformation
for amplification of the plasmid) when appropriate. Validation of
cloning and mutagenesis is determined by full-length sequencing.
The given sequence is 100% guaranteed. A report (PDF / doc) containing a
review of cloning / mutagenesis performed, sequencing results is
Expression and purification test of proteins deliverables: a detailed technical report containing an estimated expression/purification level is provided.
An analysis of expression level of recombinant protein is made by sub-cellular fractionation followed by SDS-PAGE gel migration, to estimate the expression level and integrity of the proteins.
Automated plasmid preparation deliverables: for low copy number plasmid, RoBioMol will deliver 30-40µL at 20ng/µL minimum. Concentration trueness is measured by NanoVue and guaranteed at 3.1%.
Detergent screening deliverables: a detailed technical report containing solubilisation and purification results analysed by SDS PAGE and/or Western blot. Detergent concentration trueness is guaranteed at 2.5%.
Products and generated reports are not kept after delivery.