to BioTechniques free email alert service to receive content updates.
Low Abundance Genes | Troublehsooting Forum

02/19/2013
Kristie Nybo, Ph.D.

How can I modify this plan to discover the low-abundant or non-expressed genes in the family?


The BioTechniques Molecular Biology Forums bring bench scientists together in an online community to ask for and share advice regarding laboratory problems. This week’s highlighted question comes from the DNA and General PCR Methods forum.

The BioTechniques Molecular Biology Forums bring bench scientists together in an online community to ask for and share advice regarding laboratory problems.




Q: I am working on a multi-member family of cysteine-rich peptides in plants. I found some of the genes by designing primers based on the peptide sequences and cloning the full transcripts. From the newly found transcripts, I then designed primers against conserved regions such as signal peptides and 5' untranslated regions to randomly clone homologous genes. With this approach, I am only able to find the abundant genes. How can I modify this plan to discover the low-abundant or non-expressed genes in the family?

A: For low abundant cDNA or non-expressed genes, you may have to screen a genomic library with labeled cDNA as a probe. Separate the clones based on…

A: Why don’t you try deep sequencing techniques like Illumina or 454? That will allow you to see the less abundant genes along with the abundant ones. Another idea is to prescreen the clones using the method…

To read answers to this question or to ask or advise others on this or any molecular biology topic, visit The BioTechniques Molecular Biology Forums.

Keywords:  troubleshooting forum