Equipment in Wooster

The Bio-Rad QX200 Droplet Digital PCR system provides an absolute nucleic acid quantification. The PCR reaction is partitioned into thousands of nanoliter-size oil droplets, where separate PCR reactions occur. Fluorescence of droplets with positive PCR reactions is measured and the average number of molecules per aliquot is estimated using Poisson distribution and then converted into molar concentration by dividing by the volume of the aliquot. 
A description of the QX200 ddPCR is available on the Bio-Rad website, and detailed information regarding how to set up experiments can be found in the Bio-Rad ddPCR application guide.
If you would like to use our droplet dPCR, please contact the MCIC staff.

A Bio-Rad CFX96 Touch System is a flexible real-time PCR instrument with six independently controlled thermal electric units providing even, precisely controlled temperatures at all times during the run, including ramping. It allows multiplexing of up to five targets in a single reaction (FAM, HEX, Texas Red, Cy5, and Quasar 705 dye). The CFX96 Touch includes one channel with a LED-filter photodiode combination dedicated to FRET single-plex experiments. The programmable thermal gradient feature can test eight temperatures at once to determine the optimal annealing temperature quickly. With the protocol autowriter, the user can promptly optimize reactions and generate an optimal protocol based on polymerase type, primers, and product length. 
Please refer to 'Instructions and tips for using the RT-PCR'

The Agilent Technologies 4200 TapeStation is a microfluidics electrophoresis system that provides sizing and quantitation of DNA and RNA samples and displays results in a digital format. Sizing range:
     - RNA: 100-6000nt
     - DNA: D1000: 35-1000bp; D5000: 100-5000bp
     - Genomic DNA:  200-60,000bp
More information about the tapes and sample submission can be found in this document.
To submit samples download the sample submission form and submit it electronically to the MCIC.

The Multi-Mode Synergy LX Reader Multi-Mode from BioTek enables absorbance, fluorescence, and luminescence measurements to automatically quantify many common microplate-based assays, such as nucleic acid quantification and purity assessment, protein quantification, gene expression, ELISA, BCA and Bradford protein and cell viability assays. Features:
- Continuous wavelength selection from 200nm-999nm for nucleic acid quantification (A260) and purity assessment (A260/A230 and A260/A280
-  Fluorescence filter cubes (485/535 and 630/678) allow for fluorescence-based DNA and RNA (qBit) and protein quantification and for fluorescence ELISA or gene expression assays
- Luminescence filter
- Enables micro-volume analysis using the BioTek’s Take3 plate and immediate results for up to sixteen, 2 µL samples.

The Qubit Fluorimeter 2.0 and Qubit Assay Kits (Sage Science) allow for accurate measurements of DNA or RNA concentrations by detecting target-specific fluorescence. The DNA or RNA samples are mixed with the Qubit Assay Kit reagents containing dyes that fluoresce only when bound to nucleic acids, and the concentration is read in the fluorimeter. The assay is performed at room temperature, and the signal is stable for up to 3 hours. Different Qubit Assay Kits are available for dsDNA, oligonucleotide, RNA, microRNAs and different sensitivity/concentration ranges.

The NanoDrop 2000 is a UV and visible spectrophotometer used to measure the concentration and purity of DNA, RNA and protein samples. Requires only 0.5 – 2 µl of sample and calculates sample purity ratios (260/280 nm and 260/230 nm).

The Covaris S220 Focused-Ultrasonicator patented operates at high frequencies (500KHz) and uses Adaptive Focused Acoustics Tubes and advanced electronics to deliver controlled energy precisely and accurately to sample volumes from 25 μl to 10 ml.  It's used for DNA, RNA, and chromatin shearing to specific sizes, tissue homogenization, cell lysis, compound dissolution, and particle micronization.

The Sage Science Pippin Prep is used for targeted size selection of DNA libraries for Illumina or Ion Torrent sequencing. The instrument allows to collect narrow and even fragment distributions between 100 - 1500 bp. Target sizes or ranges of sizes are entered in the software, and fractions are collected in the buffer. Up to 5 samples per gel cassette may be run, with no possibility of cross-contamination.

The Eppendorf epMotion 5075 automated liquid handling system automates routine pipetting tasks. This accurate pipetting station helps eliminate manual pipetting errors and maximizes the reproducibility of assays.
The system is set up for:
     - DNA extractions
     - Setting up PCR reactions
     - Pooling of PCR reactions from several microtiter plates
     - Preparation of Illumina libraries