STEP 1
You load the gene and the vector
GenBank, FASTA, SnapGene .dna or EMBL, from a file, by pasting the text or with an NCBI accession number. The cloning site is found by calculation, not by its name.
STEP 2
Every pair of enzymes, with its verdict
Ten checks on every pair. The ones that work are ranked; the ones that do not carry the reason and the position in writing.
623enzymes
10checks
STEP 3
Primers with two temperatures
The one for the region that anneals, which rules the first cycles, and the one for the whole oligo. Nearest neighbours with the SantaLucia (1998) parameters.
STEP 4
The gel you should see
Cut vector, PCR product and final plasmid, with the ladder chosen to suit the sizes.
Try the example
