Fragment dna by nebulization, which is like using shearing forces to create fragments. In it, you force dna in solution through a very small hole under high pressure, so fragments are made. Problem if you want to pcr these fragments: you can"t. Pcr these fragments right away because you don"t know the sequences of each end of the fragments, so you can"t make the primers. Sanger sequencing used universal primers, which were made complementary to the vector sequences. Fragment dna and ligate adaptors to ends. Ligate 2 small dna fragments (30 - 50 bases) known as either adaptors or linkers. Add some ligase to stick one adaptor to each end o the dna fragment. You need one adaptor on each end of the fragment. Select fragments with two different adaptors (for pcr amplification) Nick non-biotinylated strand to get single stranded template dna (sstdna) library. Beads are coated with streptavictin which have biotin.
