Hauser N C, Vingron M, Scheideler M, Krems B, Hellmuth K, Entian K D, Hoheisel J D
Molecular-Genetic Genome Analysis Group, Deutsches Krebsforschungszentrum, Heidelberg, Germany.
Yeast. 1998 Sep 30;14(13):1209-21. doi: 10.1002/(SICI)1097-0061(19980930)14:13<1209::AID-YEA311>3.0.CO;2-N.
Open reading frames (6116) of the budding yeast Saccharomyces cerevisiae were PCR-amplified from genomic DNA using 12,232 primers specific to the ends of the coding sequences; the success rate of amplification was 97%. PCR-products were made accessible to hybridization by being arrayed at very high density on solid support media using various robotic devices. Probes made from total RNA preparations were hybridized for the analysis of the transcriptional activity of yeast under various growth conditions and of different strains. Experimental factors that proved critical to the performance, such as different RNA isolation procedures and the assessment of hybridization results, for example, were investigated in detail. Various software tools were developed that permit convenient handling and sound analysis of the large data quantities obtained from transcriptional profiling studies. Comprehensive arrays are being distributed within the European Yeast Functional Analysis Network (EUROFAN) and beyond.
使用12232个针对编码序列末端的特异性引物，从芽殖酵母酿酒酵母的基因组DNA中PCR扩增开放阅读框（6116个）；扩增成功率为97%。通过使用各种机器人设备将PCR产物以非常高的密度排列在固体支持介质上，使其可用于杂交。用总RNA制备物制成的探针进行杂交，以分析酵母在各种生长条件下和不同菌株中的转录活性。例如，详细研究了对实验性能至关重要的实验因素，如不同的RNA分离程序和杂交结果的评估。开发了各种软件工具，以便对从转录谱研究中获得的大量数据进行方便的处理和合理的分析。综合阵列正在欧洲酵母功能分析网络（EUROFAN）及其他机构中分发。
