The Luna® Cell Ready One-Step RT-qPCR Kit provides all the necessary components for direct, dye-based RNA detection and quantitation, bypassing the need for RNA extraction and purification. The Luna Cell Ready One-Step RT-qPCR Kit is composed of: 1) the Luna Cell Ready Lysis Module (NEB #E3032S) and 2) the Luna Universal One-Step RT-qPCR Kit (NEB #E3005L).
Cell cultures are often analyzed for gene expression or treatment responses as a proxy for a living organism. Traditionally, RNA is extracted and purified from treated cells via column-based or chemical methods. Coordinating the actions of DNase I and the Luna Cell Ready Protease, the Luna Cell Ready Lysis Module offers a simple alternative workflow resulting in effective cell lysis, RNA release, and genomic DNA removal simultaneously in a 15-minute protocol. The Lysis Module includes a unique Luna Cell Ready RNA Protection Reagent that maintains RNA integrity during cell lysis. The lysis capacity spans 10 -100,000 cells in a 50 µl lysis reaction. Up to 2 µl of lysate (equivalent to RNA from 0.2 - 4,000 cells) can be transferred into 20 µl downstream RT-qPCR reactions. Similar to other Luna products, the lysis buffer includes an inert blue tracking dye for visual assistance throughout the workflow.
The Luna Universal One-Step RT-qPCR Kit is compatible with dye-based real-time quantitation of target RNA sequences via the SYBR®/FAM fluorescence channel of most real-time instruments. In the Luna Universal One-Step RT-qPCR Kit, Hot Start Taq DNA polymerase is combined with a novel Luna WarmStart Reverse Transcriptase, allowing dual control of enzyme activity via reversible, aptamer-based inhibition. The Luna Universal One-Step RT-qPCR Kit can detect and quantitate RNA transcripts in the cell lysate prepared directly from the Luna Cell Ready Lysis Module.
This table below lists cells lines that are compatible with the Luna Cell Ready Lysis module. It includes results seen using a 5-log dilution series (100,000 - 10 cells per 50 µl lysis reaction) and subsequent detection of 1 µl of lysate in a 20 µl one-step RT-qPCR reaction. For each cell line, linear results are indicated in the final column. In addition, some insect cell lines have been tested and are compatible.
Table 1: Validated Cell Lines
Cell line
Property
Species
Cell number per 50 μl lysate
A549
Adherent
H. sapiens, Lung, carcinoma
10 - 100,000
HEK293
Adherent
H. sapiens, Kidney
10 - 100,000
HeLa
Adherent
H. sapiens, Cervix, adenocarcinoma
10 - 100,000
HepG2
Adherent
H. sapiens, Liver, carcinoma
10 - 1,000
NCI-H460
Adherent
H. sapiens, Lung, carcinoma
10 - 100,000
SK-N-SH
Adherent
H. sapiens, Brain, Neuroblastoma
10 - 10,000
U2Os
Adherent
H. sapiens, Bone, osteosarcoma
10 - 100,000
Jurkat
Suspension
H. sapiens, T lymphocyte, leukemia
10 - 100,000
K-562
Suspension
H. sapiens, Lymphoblast, leukemia
10 - 100,000
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
Licenses
Nucleic acid-based aptamers for use with thermophilic DNA polymerases and reverse transcriptases are licensed exclusively by New England Biolabs, Inc. from SomaLogic, Inc. New England Biolabs, Inc. gives the Buyer/User a non-exclusive license to use the Luna® Cell Ready One-Step RT-qPCR Kit for Research Use Only (RUO). Commercial use of this product may require a license from New England Biolabs, Inc. For additional information or to inquire about commercial use, please contact busdev@neb.com.
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