Successful automation of the time-consuming and laborious steps of yeast 2-hybrid (Y2H) screening creates a high-throughput method to examine protein-protein interactions
Genetix says it has successfully automated the time-consuming and laborious steps of yeast 2-hybrid (Y2H) screening, turning this into a high-throughput method to examine protein-protein interactions.
As there are hundreds of thousands of different proteins, the number of possible interactions is vast, hence the need for rapid high-throughput screening.
Beneficially Y2H technology requires no prior knowledge of the prey proteins being tested and therefore it is possible to screen entire libraries of cDNAs encoding proteins in a high-throughput fashion.
A further advantage is that the gene sequence for the interacting proteins is contained in the clones that score positively in 2-hybrid screens, facilitating the retrieval of the genes of interest.
The Genetix Y2H screening system is available as an option on the QBot and has the capacity to process up to 100,000 Y2H interactions per day unattended.
The QBot with Y2H option automates the labour intensive steps of the Y2H experiment.
In the first step, 'prey' and 'bait' yeast colonies are arrayed at c100,000 spots per hour in high-density format onto Genetix QTrays using a 384 pin head so that they form a regular grid of yeast colonies growing on the surface of the agar.
The bait and prey colonies are then replicated on top of each other so that they can mate, and the resultant cells contain both bait and prey.
These colonies are then replica gridded onto successive selective media, to select only those cells containing positive interactions which are picked at a rate of 4000 colonies per hour onto 96 or 384-well microplates.
The Y2H option on the QBot is supplied with a powerful software package and barcode scanner for complete data tracking.
A log file in XML-format, that contains full details of all samples, is produced so that users can easily integrate the data into their own management systems.
This system has the capacity to perform Y2H screening on up to 4 x QTrays taking source bacteria from up to 72x96/384-well plates.
