Lonergan William, Whistler Toni, Vernon Suzanne D
Chronic Viral Diseases Branch, Division of Viral and Rickettsial Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
BMC Biotechnol. 2007 May 18;7:24. doi: 10.1186/1472-6750-7-24.
Several different commercial one-cycle labeling kits are available for preparation of the target for use with the Affymetrix GeneChip platform. However, there have been no evaluations of these different kits to determine if comparable results were generated. We report on the cRNA target synthesis, labeling efficiency and hybridization results using the One-Cycle Target Labeling Assay (Affymetrix), the BioArray RNA Amplification and Labeling System (Enzo Life Sciences), and the Superscript RNA Amplification System (Invitrogen Life Technologies).
The only notable difference between kits was in the yield of cRNA target synthesized during in vitro transcription, where the BioArray assay had to be repeated several times in order to have sufficient target. However, each kit resulted in comparable signal and detection calls when hybridized to the Affymetrix GeneChip.
These 3 one-cycle labeling kits produce comparable hybridization results. This provides users with several kit options and flexibility when using the Affymetrix system.
有几种不同的商业单循环标记试剂盒可用于制备与Affymetrix基因芯片平台配套使用的靶标。然而，尚未对这些不同的试剂盒进行评估以确定是否能产生可比的结果。我们报告了使用单循环靶标标记测定法（Affymetrix）、生物阵列RNA扩增和标记系统（安捷伦生命科学公司）以及超转录本RNA扩增系统（英杰生命技术公司）进行的cRNA靶标合成、标记效率和杂交结果。
各试剂盒之间唯一显著的差异在于体外转录过程中合成的cRNA靶标的产量，其中生物阵列测定法必须重复几次才能获得足够的靶标。然而，当与Affymetrix基因芯片杂交时，每个试剂盒产生的信号和检测结果相当。
这三种单循环标记试剂盒产生的杂交结果相当。这为用户在使用Affymetrix系统时提供了多种试剂盒选择和灵活性。
