LyoPrime Luna® (NEB #L4001P) 96-Well Plate Format Overview
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This video explains how to handle and use LyoPrime Luna Probe One-Step RT-qPCR Mix with UDG in 96-Well Plate Format and tips for optimal performance.
The LyoPrime Luna® Probe One-Step RT-qPCR Mix with UDG 96-Well Plate Format is lyophilized in a ‘ready-to-use’ 96-well plate format and packaged in a foil pouch with desiccant to protect the lyophilized reagent from atmospheric moisture. It is shipped and stored at room temperature prior to use, with a shelf life of 2 years.
To use the product, open the foil pouch by carefully tearing at the notch. Unseal the pouch and remove the plate. The tubes sit in a cardboard insert that provides rigidity during shipment and can be recycled.
The plate consists of twelve 8-well strip tubes that are secured in a single use yellow shell frame adaptor. The tubes are capped with optical lids and are numbered from 1 to 12 at the bottom to provide a tracking system that is independent from the yellow shell frame adaptor. The 8-well strip tubes are partially attached, allowing use of the entire plate directly or partial consumption by separating the desired number of 8-well strip tubes. Each tube contains a lyophilized cake that enables one-step probe-based detection of RNA targets in a 20 uL reaction.
The yellow shell frame adapter is compatible with ABI FAST real-time thermocyclers such as the ABI 7500 Fast or the QuantStudio® 6 and therefore may be used in full plate runs. If using the entire plate on an ABI FAST real-time instrument, remove the optical lids by pealing them off from the center edge of the plate. Continue with reaction set up.
If the plate will be used partially, individual 8-well strips can be used without the yellow shell frame. For non-ABI instruments, the removal of yellow shell frame adaptor may be required. Please refer to the protocol for recommendations on the use of the yellow shell frame adaptor in common real-time PCR instruments.
For partial consumption or for use on non-ABI instruments, remove the tubes from the yellow shelf frame grid. Set the plate on a flat surface and press down firmly on the four corners of the frame grid to release the tubes. Next, separate the desired number of 8-well strip tubes and place in a PCR rack. Any unused strip tubes should immediately be placed back in the foil pouch with the desiccant, closing the seal securely. For best results, the unused product should be consumed within one week of opening the foil pouch. Next, remove the optical lids and continue with reaction set up.
To set up the reactions, it is highly recommended to prepare an assay mix including water, primers and probes as outlined in the protocol. This will eliminate volumetric errors and ensure proper rehydration of the lyophilized mix as the liquid is dispensed into each well.
Mix the lyophilized cakes thoroughly but gently by pipetting up and down a minimum of five times. Add RNA template and recap the tubes with the optically transparent caps. Spin down the plate in a benchtop plate spinner; a plate adaptor or a carrier plate may be required. The plate is now ready for use in real time PCR.
If the tubes need to be placed back into the yellow shell frame adaptor, click the edge of each strip tube back into the adaptor.
To learn more, please visit our webpage dedicated to lyophilized products at neb.com, www.neb.com/lyoprime