Automated Affymetrix GeneChip® Sample Preparation
Sample preparation for expression studies on gene chips involves numerous steps. The procedure is time consuming and prone to errors. As a solution to this problem, HAMILTON now offers an automated system based on the MICROLAB STAR liquid handling robot. Samples from tissue are processed without manual involvement until labelled cRNA is ready for hybridization.
Tests of the protocol showed consistent cDNA and cRNA yields. Various tissue samples resulted in an average cDNA yield over 48 samples of 1.2µg in an elution volume of 25µl. The average cRNA yield over all samples was 58µg, well above the required amount of 26µg for hybridization on Affymetrix GeneChips®.
The system consists of a MICROLAB STAR liquid handling robot and integrated peripherals for incubation, agitation, analysis, etc. The complete system remains compact, due to the efficient integration of third party equipment with the built-in robotic plate handler.
The protocol is split into three methods. This allows the exclusion of bad samples with a positive/negative selection before starting the next method. All master mixes, e.g. for first and second strand cDNA synthesis, are prepared automatically by the workstation.
The flexible and scalable system allows the automation of other genomics applications such as PCR set-up, PCR reactions and PCR clean-up on the same instrument.
GeneChips® is a registered trademark of Affymetrix, Santa Clara/CA, USA.
- Automated hands-free processing starting from tissue samples and ending with labelled cRNA samples ready for hybridization
- All master mixes, e.g. for first and second strand cDNA synthesis, are prepared automatically by the robot
- Compact system due to the complete integration of third party equipment with the built-in robotic plate handler iSWAP
- Dividing the complete process into three methods gives good flexibility regarding sample processing
- Flexible system allows the automation of other genomics applications such as PCR set-up, PCR reactions and PCR clean up
- MICROLAB® STAR, 8 channels, Auto load option for barcode handling
- AGOWA® Maxisep 9600 and / or 7200 magnetic separator (AGOWA, Berlin, Germany)
- All required carriers