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.
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