The RoBioMol platform, hosted by the Pneumococcus Group of the Institut de Biologie Structurale, 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).
Contact the RoBioMol team to submit your request.After discussion and acceptance of your application, you will enter your order on RoBioNET. Acceptance of the quote (signed by both parties) will be needed prior to the beginning of the work.
Depending on the type of requested service, the gene to be mutated or expressed 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 gel electrophoresis). Samples must have security level L1 and conform to the corresponding GMO statement.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 at -80 ° C, and discarded at the end of the work unless specified otherwise on the order form.
Depending on the type of requested service Gene cloning and mutagenesis deliverables: the plasmid DNA containing the cloned or mutated gene (50ng/µL, 10µL) when appropriate and a detailed technical report. Expression and purification test of proteins deliverables: a detailed technical report containing an estimated expression/purification level. Automated plasmid preparation deliverables: plasmid DNA (80µL at 50ng/µL).
The plasmids containing the cloned or mutated genes are delivered in solution. A report (PDF doc) containing a review of cloning / mutagenesis performed, sequencing results and protein expression tests, and complete sequences of constructed vectors is transmitted by email to the customer.Plasmids products and generated reports are not kept after delivery to customers.
Validation of cloning and mutagenesis is determined by full-length sequencing. 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.
Microlab Star Hamilton for versatile molecular biology processes Nanovue GE Healthcare for matrix quality control HiGro temperature controlled tower incubator for large number of small-scale microbial cultures. CFX Connect Biorad for real time PCR and thermal shift assay processesAll these equipments are maintained and serviced rigorously.