MCB 4320C Lecture Notes - Lecture 3: Amplicon, Dideoxynucleotide
19 Jan 2019
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Document Summary
Technology is still used today but with many advances in throughput and read length. Flow cell has billions of nano-well structures. Step 1: fragment dna to desired size. The longest sequences obtained from today"s illumina technology is nearly 600 bases. In that case, 300 bases of both ends of a dna strand are sequenced. Sequencing both ends is called paired-end sequencing. When there is overlap, it is called overlapping paired-end sequencing. Step 2: ligation of adapters to both ends of dna fragments to be sequenced. Step 3: single stranded dna binds to flow cell. The flow cell has adapter sequences already bound to its surface. Dna with adapters is made single-stranded and washed across the flow cell. Each strand forms a bridge with the bridging end complementary to an adapter already on the flow cell. A full complementary strand is then made with deoxyntps and dna polymerase. Double stranded dna is denatured on the flow cell.
