Avoid multiple binding sites on the template: putative binding sites are defined as 80% of your primer sequence or 100% of last 7 bases on the 3' end aligned to another site on the template.Avoid secondary structures: hairpins, palindromic sequence, dimers, and more 3Gs or 3Cs in a stretch.What tips do you have for designing primers? The agar should not be too liquid for shipment. Please use endA- strains such as Dh4α, HB101, and XL1-Blue.
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