| 摘要 |
<p>Process for quantitative evaluation of a targeted genetic rearrangement or recombination by selective amplification of genomic DNA. Process for quantitative evaluation of a targeted genetic rearrangement or recombination comprises: (a) extracting human genomic DNA (I) from a sample; (b) amplifying segments (II) of (I) of size from a few hundred to a few tens of thousands of bases by multiplex PCR; (c) separating the amplified (II) and; (d) detecting the rearranged/recombined segments. Step (b) is done in presence of: (i) one or more primer pairs, where at least one primer hybridizes upstream of and/or at the 5'-end of, one gene (Vx) being amplified, and the other hybridizes downstream of, and/or at the 3'-end of, the other gene (Jy) being analyzed, where Vx and Jy may be involved in a genetic rearrangement, and (ii) at least one DNA polymerase that can amplify (II) (preferably of over 10 kb) and has correction activity to improve the elongation reaction. The amplification step also includes initial denaturation; cycles of denaturation, hybridization and elongation, with the elongation being for at least 10 minutes at 68-72[deg]C. Independent claims are also included for the following: (1) kit comprising the specified primers and probes, also usual buffers and reagents for performing a PCR; (2) primer for use in the new process having any of 21 sequences reproduced; and (3) detection probe for use in the new process having any of 16 sequences reproduced.</p> |
