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The association of a cassette PCR GW method with the powerful pyrosequencing technology introduced by Wanget al. [24] is of great interest since it allows mas-sive parallel sequencing of thousands and thousands of amplification fragments. A further improvement to this method was obtained by the same group, by applica-tion of a DNA barcoding strategy [23] (Fig. 1). By using cassette primers with different barcodes, authors were able to identify more than 160 000 integration sites for lentiviral and gamma-retroviral vectors in sev-eral tissue samples from mice. A similar approach was reported for the analysis of maize transposons by Liu et al. [27] in the blocked digestion–ligation–amplifica-tion (blocked DLA) method. The method adopts a sin-gle-strand adaptor provided with a 3 ¢ -terminus annealable with 3¢ -overhangs of genomic restriction fragments obtained by NspI(RCATG• Y) digestion. After the first step, the library fragments are blocked in 3¢ -termini by addition of dideoxynucleotide and then amplified with sequence specific and adaptor primers. In the so-called MuClone strategy 更多：https://www.bmcx.com/ , DLA is adapted to the identification of gene sequences flanking the highly active maize mutator ( Mu) transposon, by using nested gene specific primers ending, downstream of the CATG NspIsequence, with all the possible three nucleotide tags starting with a C or T (so to be com-patible with the completeNspIsite). By combining the MuClone strategy with pyrosequencing technology the authors obtained about 965 000 reads [60].