IDT CUSTOM OLIGOS:

**New** Order custom oligos through IDT here

To order oligos from IDT:
1. Click the link above or copy and paste the following address in your browser. https://www.idtdna.com/njms
2. Create an account (account will be confirmed by MRF).
3. Log in and place order.
4. Pick up products in MSB F503 or request delivery to ICPH and Cancer Center.

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FRAGMENT ANALYSIS: Fluorescently labeled fragments are detected using the Applied Biosystems 3130XL Genetic Analyzer and then interpreted using the GeneMapper analysis software. Five different colored fluorescent dyes can be detected in one sample. The analysis software uses the size standard to create a standard curve for each lane and then determines the length of each dye-labeled fragment by comparing it with the standard curve for that specific lane.

OLIGO SYNTHESIS : Oligonucleotides are synthesized on two Applied Biosystems 394 DNA/RNA Synthesizers. Oligonucleotides can be produced up to 100 nucleotides in length at various synthesis scales which produce up to milligram quantities of high-quality oligonucleotides for use as primers, probes or other applications. The synthetic oligonucleotides can be purified by Oligonucleotide Purification Cartridges(OPC). Specialized nucleotides can be added to the sequences as well as labeling of the oligos with fluorescent or other derivatives.

REAL TIME PCR :The facility houses two Real Time PCR systems.The LightCycler® 2.0 Instrument (Roche) allows you to monitor amplification of a PCR product simultaneously, in real-time and online, with six different detection channels. The LightCycler® Software 4.05 collects fluorescence data from all six channels at each cycle and displays it immediately.

The Applied Biosystems 7500 Real Time PCR Systems uses fluorescent-based PCR chemistries to provide quantitative detection of nucleic acid sequences using real-time analysis and qualitative detection of nucleic acid sequences using end-point and association-curve analysis.

PEPTIDE SYSNTHESIS : Peptides are synthesized on the Applied Biosystems model 433 peptide synthesizer which is capable of producing milligram quantities of peptides up to twenty five amino acids. After cleavage and deprotection the peptides can be purified by size exclusion chromatography or by HPLC.

PROTEIN SEQUENCING: Purified proteins are sequenced using the Applied Biosystems Procise protein sequencer with results up to fifteen amino acids per sequence. Proteins are prepared for sequencing either in solution or transferred from acrylamide gels to PVDF membranes. Results from the sequencing reactions are analyzed using the related software and the results presented in both graphic and tabular form.

HPLC PURIFICATION AND ANALYSIS: HPLC purifications noted above and analysis of a wide range of samples is performed with the Dionex DX500 HPLC system. This system is controlled through a set of operational and analytical software programs in the PeakNet Chromatography Workstation package that allows for detailed analysis of the data collected on the AD20 absorbance detector which will monitor in the visible as well as UV wavelengths.

IMAGING: The facility houses two Typhoon imagers, a fluorimager FSI (Molecular Dynamics) for scanning of filters, gels, microtiter plates, etc. that are labeled with fluorescent tags, two phosphorimagers (SI and 45SI, Molecular Dynamics) for scanning radioactively labeled materials in gels or filters and a densitometer. Each of these scanners is supported by software that allows both quantitation (ImageQuaNT) and size determination (FragmeNT) of scanned images.

AMAXA SYSTEM: The Nucleofector Device- The Nucleofector technology is a highly efficient non-viral method for transfection. It is based on two components: the Nucleofector Device that delivers unique electrical parameters, and Nucleofector Kits that contain cell-type-specific Nucleofector Solutions.

LUMINEX SYSTEM: Luminex® xMAP® technology offers ideal speed and sensitivity for performing multiplexed cytokine measurements. Luminex’s approach offers comparable sensitivity to traditional ELISA-based systems, but with additional advantages including extended dynamic range and smaller sample size.Luminex 100 IS system is developed based on xMAP® technology and it is a flexible analyzer based on the principles of flow cytometry. The Luminex 100 IS System is the combination of three core xMAP technologies. The first is xMAP microspheres, a family of 100 fluorescently dyed 5.6 micron-sized polystyrene microspheres that act as both the identifier and the solid surface to build the assay. The second is a flow cytometry-based instrument, the Luminex 100, which integrates key xMAP detection components such as lasers, optics, advanced fluidics and high-speed digital signal processors.

5PRIME RNA PURIFICATION SYSTEM:

The PerfectPure RNA 96 CellVac Kit provides the components and procedures necessary for high-throughput purification of total RNA from cultured cells.Vacuum processing is used throughout the entire purification resulting in RNA that is essentially free of genomic DNA, protein, and other enzymatic inhibitors.High-quality RNA is required in RNA analysis procedures such as RT-PCR,microarray analyses, northern blots, dot blots, in vitro translations, ribonuclease protection assays, and other downstream analyses. The PerfectPure RNA 96 Cell Kit comes complete with pre-mixed reagents and provides an efficient method of extracting RNA from 192 samples in approx 2 hours. The RNA purification process utilizes novel, proprietary reagent formulations that eliminate the use of hazardous chaotropic reagents.

 The PerfectPure RNA 96 CellVac Kit (2302540) uses a vacuum-based method to purify high quality RNA from cultured cells and the PerfectPure RNA 96 CellCS Kit (2302530) uses centrifugation process to extract high quality RNA . Total RNA is purified with the PerfectPure RNA 96 CellVac Kit by first adding a detergent/salt solution to the cultured cell samples to lyse and homogenize the cells and eliminate endogenous RNase activity. Homogenization disrupts the cell membranes releasing RNA into the lysis solution. Next, the lysates are applied to the plate to bind the RNA and wash away proteins, DNA, and other contaminants. Residual DNA is then removed by an on-plate DNase treatment. Finally, purified RNA is eluted with DEPC-treated water.