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[Isolating the ends of yeast artificial chromosome by inverse polymerase chain reaction]
1Department of Medical Genetics, Shandong Medical University, Jinan, Shandong, 250012 P. R. China. Yaoqing@hotmail.com
Abstract
Objective:
To establish a highly efficient method for isolating yeast artificial chromosome(YAC) ends.
Methods:
Based on the sequence of TAC vector, the frequent cutting enzymes were used to cleave within the vector and the genomic insert to generate relatively small fragments,which were ligated and subsequently amplified using vector primers that are in inverse orientation. The PCR products were purified and sequenced. RESULTS 8 YAC termini were isolated from YAC 776E2,964C5,11D8 and 8D1.
Conclusion:
The results suggest that inverse PCR be an efficient method for isolating YAC termini.