Comparing Reagents Used on Applied Biosystem's 3730xl

摘要

Reagents are an integral part of sequencing on Applied Biosystems (ABI) 3730xl. They allow the samples to move through the machine with ease, providing clarity and quality results. But are all reagents created equal? We compared 5x Sequencing Buffer, Dye Terminator cycle sequencing reagent, 10x Running Buffer, and Sequencing Polymer from Edge Biosystems (Edge), ABI, and McLab in order to determine how well each of them performed by evaluating array images, raw fluorescence signal, read clarity, peak uniformity, and sequencing start point. Twelve PCR products and four plasmid samples underwent cycle sequencing using Dye Terminator cycle sequencing reagent and Sequencing Buffer from each of the three companies: Edge, ABI, and McLab. Each of the three groups of 16 samples were sequenced using Edge Running Buffer and Polymer, ABI Running Buffer and Polymer, and McLab Running Buffer and Polymer. Additionally, each group of 16 samples were also sequenced using each combination of Edge, ABI, and McLab Running Buffer with Edge, ABI, and McLab Polymer. For example, 16 samples prepared with Edge Dye Terminator and Sequencing Buffer were sequenced using: Edge Running Buffer/ABI Polymer, Edge Running Buffer/McLab Polymer, ABI Running Buffer/Edge Polymer, ABI Running Buffer/McLab Polymer, McLab Running Buffer/ABI Polymer, and McLab Running Buffer/Edge Polymer, and so on. After testing, we looked at array images for background clarity. We compared traces for signal strength and read consistency, and we viewed the sample start point to see when each sequence began. Overall, we determined that all of the reagents were comparable, with Edge and ABI performing slightly better in signal strength, read clarity, and earlier start points.