2. Select a
gene from
your viral
genome
Press Search
Select Nucleotide from list
Put Put gene name in NCBI search bar
• use proper spellings
Go Go to NCBI at
https://www.ncbi.nlm.nih.gov/
Searc
h
Gene name
3.
4.
5. Sequence selection
Select at least 5 sequences
• Open word file and paste sequences in FASTA format
Go to Clustal Omega at
https://www.ebi.ac.uk/Tools/msa/clustalo/
• Select DNA
• Put all sequences in box
• Submit
Result
Select sequence with least mutations
8. Primer 3 plus
Go to webpage at https://www.primer3plus.com/
Put sequence in box
Adjust parameters
• Primer size = 18-22
• Tm = 55-65
• GC % = 40-80
• Product size = 150-350
Select pick primers