Go direct from cells to RNA quantitation without purification
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 |
The following reagents are supplied with this product:
NEB # | Component Name | Component # | Stored at (°C) | Amount | Concentration |
E3030S | -20 |
Luna Cell Ready Lysis Buffer | B8036AVIAL | -20 | 2 x 1.4 ml | 2 X | |
Luna Cell Ready RNA Protection Reagent | M0469AVIAL | -20 | 1 x 0.25 ml | 25 X | |
Luna Cell Ready Protease | M0473AVIAL | -20 | 1 x 0.2 ml | 25 X | |
Luna Cell Ready Stop Solution | B8037AVIAL | -20 | 1 x 0.6 ml | 10 X | |
DNase I (RNase-free) |
M0474AVIAL | -20 | 1 x 0.5 ml | 10 X | |
Luna® WarmStart® RT Enzyme Mix | M3002LVIAL | -20 | 1 x 0.5 ml | 20 X | |
Luna® Universal One-Step Reaction Mix | M3005SVIAL | -20 | 5 x 1 ml | 2 X | |
Nuclease-free Water | B1502AVIAL | -20 | 3 x 1.5 ml |
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.
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.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.