TAILIEUCHUNG - Báo cáo khoa học: Modified PCR methods for 3¢ end amplification from serial analysis of gene expression (SAGE) tags

Serial analysis of gene expression (SAGE) is a powerful technique to study gene expression at the genome level. However, a disadvantage of the short-ness of SAGE tags is that it prevents further study of SAGE library data, thus limiting extensive application of the SAGE method in gene expression studies. However, this problem can be solved by extension of the SAGE tags to 3¢ cDNAs. | ỊFEBS Journal REVIEW ARTICLE Modified PCR methods for 3 end amplification from serial analysis of gene expression SAGE tags Wang-Jie Xu1 Zhao-Xia Wang1 and Zhong-Dong Qiao1 2 1 College of Life Science and Technology Bio-X Research Center Key Laboratory of DevelopmentalGenetics and Neuropsychiatric Diseases Ministry of Education Shanghai Jiao Tong University China 2 Shanghai Institute of MedicalGenetics Shanghai Jiao Tong University China Keywords 3 longer fragment cDNA generation of longer cDNA fragments from SAGE tags for gene identification GLGI high-throughput methods mRNA rapid RT-PCR analysis of unknown SAGE tags RAST-PCR reverse SAGE rSAGE serial analysis of gene expression SAGE tag two-step analysis of unknown SAGE tags TSAT-PCR Correspondence Z. Qiao College of Life Science and Technology Shanghai Jiao Tong University 800 Dongchuan Road 200240 Shanghai China Fax 86 21 5474 7330 Tel 86 21 3420 4925 E-mail zdqiao@ Serial analysis of gene expression SAGE is a powerful technique to study gene expression at the genome level. However a disadvantage of the shortness of SAGE tags is that it prevents further study of SAGE library data thus limiting extensive application of the SAGE method in gene expression studies. However this problem can be solved by extension of the SAGE tags to 3 cDNAs. Therefore several methods based on PCR have been developed to generate a 3 longer fragment cDNA corresponding to a SAGE tag. The list of modified methods is extensive and includes rapid RT-PCR analysis of unknown SAGE tags RAST-PCR generation of longer cDNA fragments from SAGE tags for gene identification GLGI a high-throughput GLGI procedure reverse SAGE rSAGE two-step analysis of unknown SAGE tags TSAT-PCR etc. These procedures are constantly being updated because they have characteristics and advantages that can be shared. Development of these methods has promoted the widespread use of the SAGE technique and has accelerated the speed of studies of large-scale gene

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