Real-time PCR Arrays
Nick A. Saunders
from: Real-Time PCR: Advanced Technologies and Applications (Edited by: Nick A. Saunders and Martin A. Lee). Caister Academic Press, U.K. (2013)
Real-time PCR arrays are tools that allow convenient testing of samples in many assays concurrently, parallel testing of many samples or testing of multiple samples and targets simultaneously. It is desirable to standardise and automate primer and probe selection due to the large number of assays that must be designed. Furthermore, it is useful to use probe selection techniques that increase the robustness of the individual assays since this will increase the level of compatibility between the assays and decrease the complexity of interpretation of the outputs. A simple approach to creating real-time PCR arrays is to use microtitre plates which currently have capacities of 96, 384 or 1536 features. Such arrays can be populated with user designed assays or with tests selected form a menu of over one million that are commercially available. A primary application of such arrays has been to verify gene expression data obtained using hybridisation. Cramming additional features into a device of manageable scale has led to the introduction of nanolitre volume arrays that diverge from the microtitre plate pattern. Several thousand different reactions can now be included in a single real-time PCR array. The reduction in scale also has advantages in terms of the volumes of materials required. As real-time arrays are miniaturised the number of pipetting steps required increases and it is often necessary to pre-configure them commercially leading to relative inflexibility. This limitation has prompted the development of arrays that include microfluidic channels and valves. These 'chips' can be loaded via relatively few liquid handling steps to create custom applications read more ...