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文献和实验Primer Design and Primer-Directed Sequencing
The dideoxy chain termination DNA sequencing procedure introduced in 1977 (1 ) has the advantage of being fast, simple to perform, and very accurate. Therefore, it became the method of choice to obtain several hundred bases of sequence
Primer Design（Manual and Automated Primer Design for SEQ and PHY Gaps）
. GC Clamp Avoid runs of identical nucleotides (e.g.ATCGACCCCCTAGAC). Similar Tm (+/-2℃) for primers in same reaction set. Unique to the template. Designing Primers for Sequencing Gaps
quantitation is critical for successful sequencing reactions. The preferred method for quantitation is by gel electrophoresis with a DNA mass ladder standard.1. Recommended amount of template and primer to use in a cycle sequencing reaction: Reaction Set
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