I am looking for target gene forward and reverse primers form published literature but it available information is limited because it lacks number of nucleotides/bp and qPCR protocol for each target gene.. available literature only intended the name of target gene and primer sequence and gene accession number as it is shown in picture. While qPCR requires qPCR protocol for each target gene for particular primer and number of nucleotides/bp for primer. However, through gene accession number i have found the target gene sequence of the origin segment of the gene. Can this sequence be used to design forward and reverse primer if yes then please share a link or source where i can use the gene sequence to design primers online?
Thanks in Advance :)
There are many online tools for primers design, for conventional PCR you can use the NCBI tool: https://www.ncbi.nlm.nih.gov/tools/primer-blast/
For qPCR (TaqMan-Real-time PCR), you can use:
https://www.genscript.com/tools/real-time-pcr-taqman-primer-design-tool
https://eurofinsgenomics.eu/en/ecom/tools/qpcr-assay-design/
Do you want primers for conventional "end point" PCR or for qPCR?
I also vote for the linked NCBI primer tool - it's really easy to use.
Also, you should be able to determine the exact product size for the published primer pairs by aligning the primers to the sequence (use the spliced version/transcript, not gDNA).
Katie A Burnette I want to do qPCR, but number of primer nucleotides are missing in literature.. but gene accession number is available.. through gene accession number a primer can be designed?
Not sure what you mean by "number of primer nucleotides". Do you mean the expected product size? Or the location of the primers in the gene of interest? Either way, if they give you the sequence of the primers, you can align them to the target gene either using an alignment tool or by hand and determine the expected product size too.
You could always email the corresponding author and ask for the exact primer sequences, assuming that they are not listed in the methods or supplemental figures.
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