Array-based oligonucleotide synthesis can be used to generate primer pools for use in MPE-seq
Obtaining adequate amounts of primer pools for MPE-seq from cost-effective array based oligonucleotide synthesis can be achieved in four steps: (1) PCR amplification of the oligonucleotide synthesis pool using a 5’ blocked sense primer and a biotinylated antisense primer. (2) Restriction digestion to cleave off the PCR primer handle. (3) Lambda exonuclease digestion of free 5’ ends. (4) Streptavidin purification of biotinylated PCR handle. The unbound fraction is the desired primer pool product.