Recipient Organization
UNIVERSITY OF TEXAS AT TYLER, THE
3900 UNIVERSITY BLVD
TYLER,TX 757990001
Performing Department
College of Arts & Sciences
Non Technical Summary
It is becoming increasingly clear that differences among individuals of the species and of different species arise largely from differences in the expression patterns of genes. For example, although humans and chimpanzees share many of the same genes, the differences between these two species are stark and clear. Studies have shown such stark differences can best be explained by differences in gene expression patterns. The proposed equipment will be used to investigate gene expression differences in a variety of animal and plant systems to understand both fundamental processes as well as phenomena that can be exploited for pest management and food production.
Animal Health Component
40%
Research Effort Categories
Basic
40%
Applied
40%
Developmental
20%
Goals / Objectives
This proposal is a request for the purchase of a high throughput quantitative, real time PCR (qRT-PCR) system that will enable multiple users in the Department of Biology, The University of Texas at Tyler, to undertake gene expression analyses on thousands of genes simultaneously. Specifically, we propose to purchase a fully integrated CFX-384 Real-Time PCR Detection System with CFX Automation System (Bio-Rad) priced at $98,530 of which we request 50% ($49,265) from USDA; non-federal matching funds are 50%. The equipment will enhance research infrastructure at UT Tyler and enable Biology faculty to generate preliminary data more efficiently and compete for grant funds more effectively.
Project Methods
Understanding biological complexity arising from patterns of gene expression requires accurate and precise measurement of RNA levels across large numbers of genes simultaneously to identify gene regulatory networks. High throughput sequencing facilitates the assay of hundreds or thousands of genes simultaneously. The detection chemistry of qRT-PCR procedures is based on monitoring amplification products through either binding of dsDNA using ethidium bromide or SYBR green or target specific hybridization to single stranded DNA using molecular beacons or hydrolysis probe assays. Until recently, throughput has been limited to 384 reactions per run for individual laboratories (e.g., LightCycler 480 (Roche), ABI PRISM 7900 HT system (Applied Biosystems); CFX PCR Detection System expands the capacity to 7680 samples. Each system has its advantages and disadvantages but they all perform equally well in terms of sensitivity and robustness of the results. They vary in sample capacity, reagent use, and time required to complete the analysis.The CFX PCR detection system and the CFX automation system are equipped with a plate handler and a capacity to load twenty 384-well plates for a total of 7680 samples at a time which facilitates workflow automation, generation of large volume of data, and rapid analysis of the data with built-in software. The software enables the investigator to define PCR protocols for multiple experiments at once allowing for different protocols for each plate. This is a useful feature because the optimal PCR conditions are often gene-specific. The qRT-PCR software for the CFX system includes various components that allow the investigator to meet the MIQE guidelines for publishing qRT-PCR data. Finally, the system also facilitates stand-alone operation or operation with multiple plates very easily.