Culture Collections

Mouse cDNA now available from ECACC

PCR-ready cell line cDNA: a transcriptome tool for cell line selection

 

Mouse cDNA - HOMEPAGE

 

One common question which regularly appears in the ECACC technical enquiries box is: “Do you have a cell line that expresses our gene of interest?”  Sometimes a literature search provides the answer but more often experimental investigation is needed.  Messenger RNA (mRNA) analysis can assess the gene expression of cell lines. Typically a researcher would isolate total RNA from the cell lines under review and convert this to complimentary DNA (cDNA) using reverse transcription.

This cDNA snapshot can be tested for the expression of the genes of interest using a PCR reaction with primers specific for certain biomarkers.  The technique does not confirm expression of a functional protein but it is relatively quick, inexpensive and allows researchers to potentially identify a more relevant cell culture model from the hundreds of candidates available from a culture collection without relying on the literature.

In ECACC we make this process easier by providing high quality RNA from all our cell lines; for many PCR-ready cDNA is also available. In the past ECACC manufactured and qualified cDNA from human cell lines only, but now, using QRT PCR and species specific reference gene primers, we also provide cDNA from mouse cell lines (Table 1).

 

 

 

Target

Sample

Human GAPDH

Mouse CANX

Mouse SDHA

HeLa (Human) cDNA

R Amplification

X   No Amplification

X   No Amplification

K38 Keratinocyte (Mouse) cDNA

X   No Amplification

R   Amplification

R   Amplification

HeLa (Human) RNA*

X   No Amplification

X   No Amplification

X   No Amplification

K38 Keratinocyte (Mouse) RNA*

X   No Amplification

X   No Amplification

X   No Amplification

Table 1: QC of mouse cDNA generated from K38 Keratinocyte (mouse) RNA

*to confirm absence of genomic DNA contamination in the RNA pre-reverse transcription

 

 

We have completed a successful trial study with the European Molecular Biology Laboratory (EMBL) in Italy who enquired if the mouse Keratinocyte cell line K38 was known to express the interleukin 31receptor (IL31RA) and oncostatin M receptor (OSMR). ECACC provided cDNA from the cells to allow EMBL to confirm gene expression by QRT PCR giving them confidence in using the cell line in their studies (see figure 1 below).

 

Mouse cDNA - CONTENT

Figure 1: QRT PCR analysis of K38 cDNA from ECACC demonstrating expression of IL31RA and OSMR using Ubiquitin as a reference gene (data courtesy Dr Linda Nocchi, EMBL)

Click here to enlarge figure 1

Following our success in supplying mouse cDNA we hope to be able to provide cDNA from many other species.                       

 

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