Molecular Quantification Laboratory

The MQL is equipped with an Applied Biosystems (ViiA7) quantitative real-time polymerase chain reaction (qRT-PCR) machine. This instrument is available for accurate and sensitive high-throughput measurements of gene expression. This instrument is also equipped with Protein Thermal Shift technology for analysis of protein stability and binding interactions. This new technology can be used for high-throughput screening of proteins to identify ligands, mutations/modifications, or buffer conditions to increase melting temperature and the relative stability of proteins. This application can also screen antibody–ligand binding as part of antibody development, or be used to assess protein preps for contaminants or aggregated protein.

MQL Rules and Guidelines

ViiA7 Applications Training

Bio-Rad Consumables recommended for AB Viaa7

MQL Rules and Guidelines

Greetings Molecular Quantification Laboratory (MQL) users (i.e. qRT-PCR people)

The Applied Biosystems (ViiA7) is currently in room 2.276 (BSB) but will moves to room 344 (IRP) in January of 2020. I will give one key to each participating laboratory. The use of USB drives will not be allowed on the instrument or on the computer that runs the instrument. You only need to give the program your e-mail address and the data will be sent to you upon completion. This will require you to load the analysis software on a designated computer for your laboratory. A license free version of the software is available. Use this link to get the "QuantStudio™ Software V1.1—for QuantStudio™ 6 and 7 Flex and ViiA™ 7 Real-Time PCR Systems" software. Mac users will have to find a way to run the PC software for the ViiA7. Alternatively, an Excel file can be generated on the MQL computer and you can e-mail it to yourself.

Whenever you want to start, just contact me and set up a time for training. We have an online signup calendar to manage usage and a paper sign in/out sheet in the room. There will be no charges for using the MQL at this time but that may change in following years.

Some important points are:

• Do not use 200ul 96-well plates! The block is for 100ul fast-run plates. If you put the 200ul plate into the machine it will be crushed and potentially damage the instrument. Initially I have the proper plates for you to use (MicroAmp Fast 96-well Optical Reaction Plates, Life Tech. #4346906).
• No USB drives even for Mac users.
• No writing on the well cover of your plate. This will be vaporized by the heated cover and will obscure the fluorescent light source.
• Do not touch the outside of your wells (with or without gloves). This will result in residue collecting inside the block well and will add to signal variability.
• Do not use benchtop cloth diapers when preparing your plate as the lint and other fibers will bake into the block wells and obscure data collection.
• Do not put the plate on ice. Salts and impurities in the water will precipitate in the block well and obscure data collection.
• Please use designated plate holders to prevent lint or residue from contaminating the outsides of your plate (e.g. MicroAmp 96-well Support base, Life Tech, cat# 4379590).
• You must sign up before using the instrument and please only sign up for the time you will need to use the instrument (within reason).
• Please remove your plate when your analysis is done.

The use of the MQL is a privilege and repeat offenders may have their access revoked.

Thanks,

Bob Marr

MQL Contact Information