Bentley-Hewitt Kerry L, Hedderley Duncan I, Monro John, Martell Sheridan, Smith Hannah, Mishra Suman
The New Zealand Institute for Plant & Food Research Ltd, Palmerston North 4410, New Zealand.
The New Zealand Institute for Plant & Food Research Ltd, Palmerston North 4410, New Zealand.
N Biotechnol. 2016 May 25;33(3):380-6. doi: 10.1016/j.nbt.2016.01.002. Epub 2016 Jan 25.
Experimental methods are constantly being improved by new technology. Recently a new technology, NanoString®, has been introduced to the market for the analysis of gene expression. Our experiments used adipose and liver samples collected from a rat feeding trial to explore gene expression changes resulting from a diet of 7.5% seaweed. Both quantitative real-time polymerase chain reaction (qPCR) and NanoString methods were employed to look at expression of genes related to fat and glucose metabolism and this paper compares results from both methods. We conclude that NanoString offers a valuable alternative to qPCR and our data suggest that results are more accurate because of the reduced sample handling and direct quantification of gene copy number without the need for enzymatic amplification. However, we have highlighted a potential challenge for both methods, which needs to be addressed when designing primers or probes. We suggest a literature search for known splice variants of a particular gene to be completed so that primers or probes can be designed that do not span exons which may be affected by alternative gene sequences.
实验方法不断因新技术而得到改进。最近,一种名为NanoString®的新技术已投放市场用于基因表达分析。我们的实验使用了从大鼠喂养试验中收集的脂肪和肝脏样本,以探究7.5%海藻饮食导致的基因表达变化。我们采用了定量实时聚合酶链反应(qPCR)和NanoString方法来研究与脂肪和葡萄糖代谢相关的基因表达,本文对这两种方法的结果进行了比较。我们得出结论,NanoString为qPCR提供了一种有价值的替代方法,并且我们的数据表明,由于减少了样本处理以及无需酶促扩增即可直接定量基因拷贝数,结果更加准确。然而,我们强调了这两种方法都存在的一个潜在挑战,在设计引物或探针时需要加以解决。我们建议完成对特定基因已知剪接变体的文献检索,以便设计出不跨越可能受可变基因序列影响的外显子的引物或探针。