Primer Tails and Adapter Sequences
A 5' extension carrying a restriction site, an overlap for Gibson assembly, a barcode or a sequencing adapter is added because only the 3' portion needs to match the template. The tail is excluded from annealing Tm for early cycles but becomes part of the template afterward, so later cycles anneal at a higher effective Tm.
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A 5' extension carrying a restriction site, an overlap for Gibson assembly, a barcode or a sequencing adapter is added because only the 3' portion needs to match the template. The tail is excluded from annealing Tm for early cycles but becomes part of the template afterward, so later cycles anneal at a higher effective Tm.
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