FAQ: What cell lines are compatible with the Luna Cell Ready Lysis Module (NEB #E3032)?

Numerous cell lines can be used for direct RNA detection using the Luna Cell Ready Lysis module paired with Luna Universal One-Step RT-qPCR kit or Luna Universal Probe One-Step RT-qPCR kit. However, some cell lines may contain high levels of inhibitors that may affect cell lysis or the RT-qPCR reaction. Culture medium and drug/RNAi/CRISPR treatment can also affect cell growth status and levels of inhibitors.

The table below highlights the results observed with various mammalian cell lines 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 experiment. For each cell line, linear results are indicated in the final column of the table below. In addition to these mammalian cell lines, the kit is also compatible with insect cell lines. For cell lines not listed below, it is recommended to perform a pilot test for the optimal cell input numbers for lysis as well as the amount of lysate that can be carried into the RT-qPCR.

Cell lines

Property

Species

Cell number per 50 μl lysate

HeLa

Adherent

Hs. Cervix, adenocarcinoma

10 - 100,000

A549

Adherent

Hs. Lung, carcinoma

10 - 100,000

Jurkat

Suspension

Hs. T lymphocyte, leukemia

10 - 100,000

HEK293

Adherent

Hs. Kidney

10 - 100,000

U2Os

Adherent

Hs. Bone, osteosarcoma

10 - 100,000

HepG2

Adherent

Hs. Liver, carcinoma

10 - 1,000

K-562

Suspension

Hs. Lymphoblast, leukemia

10 - 100,000

NCI-H460

Adherent

Hs. Lung, carcinoma

10 - 100,000

SK-N-SH

Adherent

Hs. Brain, Neuroblastoma

10 - 10,000