Production of shotgun libraries using random amplification

In the following report, thermal cycling coupled with random 10-mers as pri mers was used to construct randomly amplified shotgun libraries (RASLs). Th is approach allowed shotgun libraries to be constructed from nanogram quant ities of input DNA. RASLs contained inserts from throughout a target genome in an unbiased fashion and did not appear to contain chimeric sequences. T his protocol should be useful for shotgun sequencing the genomes of uncultu rable organisms and rapidly producing shotgun libraries from cosmids, fosmi ds, yeast artificial chromosomes (YACs), and bacterial artificial chromosom es.