Things To Consider When Ordering Published Primer Sequences

When you see a set of primers in the materials and methods section of a journal publication, it is tempting to order the sequences provided. Before doing so, it is good practice to check the primer sequences to make sure that they are suitable for your needs. In other words, make sure the primer sequences published are suitable for your needs in regards to:

* Cell type – some cell types may have little to no expression of your GOI.

* Tissue type – as with cell type, there may be little to no expression of your GOI.

* Species and percent homology if the species are different.

* Whether the region targeted in the transcript will give you the expected product size when PCR products are run on a gel.

* Primer direction – sometimes the primer sequences provided in a journal article may not be in the correct format for ordering. For instance, if a reverse sequence rather than the reverse complement sequence is provided in the publication, you will have serious issues when you start experimenting with the primers.

* Purpose of the primer – it is important to see what experiments the sequences published were actually used for. A primer set used for expression cloning will not be suitable for real-time PCR.

* The primer sequence published – you want to make sure that the sequences provided are accurate so check by blasting the sequence. It is not uncommon for authors to make typos when entering in their sequences during manuscript preparation.