Product Class: Kit

Luna® Cell Ready Lysis Module

Product Introduction

  • Coordinated cell lysis, RNA release, and genomic DNA removal in a fast 15-minute protocol
  • Effective cell lysis preparation from 10 to 100,000 cells across numerous cell lines
  • Minimal sample loss compared to alternative RNA extraction methods, especially at low cell input
  • Compatible with high-throughput applications: cell lysis can take place at room temperature
  • Optimal performance when paired with Luna Universal One-Step RT-qPCR kits: NEB #E3005 (dye-based) or NEB #E3006 (probe-based)
  • Complete lysis and one-step RT-qPCR workflow available: NEB #E3030 (dye-based) or NEB #E3031 (probe-based) 
 
Bioz Badge Exists : True
Catalog # Size Concentration
E3032S 100 reactions

Product Information

Description


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. The Luna Cell Ready Lysis Module is a fast, convenient, and sensitive alternative for evaluating RNA expression levels without performing traditional RNA extraction steps. The module coordinates cell lysis, RNA release, and genomic DNA removal simultaneously, resulting in cell lysate ready for direct RT-qPCR analysis with Luna Universal One-Step RT-qPCR kits. Similar to other Luna products, the lysis buffer includes an inert blue tracking dye for visual assistance throughout the workflow.

This table lists cell 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) 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


Figure 1: Luna Cell Ready Workflow



Coordinating the actions of DNase I and the Luna Cell Ready Protease, the Luna Cell Ready Lysis Module offers a simple workflow resulting in effective cell lysis, RNA release, and genomic DNA removal simultaneously in a 15-minute protocol. Up to 2 µl lysate (equivalent to RNA from 0.2 - 4,000 cells) can be transferred into 20 µl downstream RT-qPCR reactions.


 
This product is related to the following categories:
Luna® qPCR & RT-qPCR,
PCR, qPCR & Amplification Technologies,
This product can be used in the following applications:
Probe-based qPCR & RT-qPCR,
qPCR & RT-qPCR,
Two-Step RT-qPCR,
DNA Amplification, PCR & qPCR

Kit Components

Kit Components

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • NEB # Stored at (°C)
    E3032S -20
    Component Name Component # Stored at (°C) Amount Concentration
    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

Properties & Usage

Materials Required but not Supplied

  • Phosphate-buffered saline (PBS)
  • Cells
  • Eppendorf tubes
  • PCR strip tubes
  • PCR plates
  • Pipettors and pipette tips (to minimize cross contamination, filter tips should be used)

FAQs & Troubleshooting

FAQs

  1. What cell lines are compatible with the Luna Cell Ready Lysis Module (NEB #E3032)?
  2. How many cells should I use for reliable detection?
  3. How many cells should I seed into cell culture plates to achieve optimal detection?
  4. Can cryopreserved cell lines or pelleted cells stored at -80°C be used with this kit? 
  5. How can lysis efficiency be determined?
  6. Why is it important to evaluate RNA damage or inhibition during cell lysis and one-step RT-qPCR detection?
  7. What is the “Luna Cell Ready RNA Protection Reagent” and when should it be used?
  8. Should I include a No-RT control reaction?
  9. Will the blue tracking dye interfere with detection?
  10. Is cell washing prior to lysis necessary?
  11. Which lysis format should I use?
  12. What protocol should be used for cell lysis? Can cells be lysed at room temperature or using a short protocol?
  13. Can I use the cell lysates generated from the Luna Cell Ready Lysis Module for two-step RT-qPCR? 
  14. When using cell lysates generated from Luna Cell Ready Lysis Module, can RT-qPCR experiments be set up at room temperature?
  15. How much cell lysate should I use as input with Luna Cell Ready One-Step RT-PCR?
  16. Why does the Luna Cell Ready One-Step RT-qPCR Kit contain more RT-qPCR reactions than lysis reactions?
  17. If either the Luna Cell Ready Lysis Module or Luna Universal One-Step RT-qPCR Kit runs out, can I order them separately?
  18. How can I obtain more technical help related to Luna Universal One-Step RT-qPCR Kit?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

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.

Licenses

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.

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc. For more information about commercial rights, please email us atgbd@neb.com While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications

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