筛选通过使用扩增RNA的差异显示获得的差异表达cDNA克隆。
Screening differentially expressed cDNA clones obtained by differential display using amplified RNA.
作者信息
Poirier G M, Pyati J, Wan J S, Erlander M G
机构信息
R.W. Johnson Pharmaceutical Research Institute, San Diego, CA 92121, USA.
出版信息
Nucleic Acids Res. 1997 Feb 15;25(4):913-4. doi: 10.1093/nar/25.4.913.
Abstract
The major obstacle of differential display is not the technique itself but rather the post-differential display issueof discriminating between false positives and the truly differentially expressed mRNAs. This process is arduous and requires large amounts of RNA. We present and validate a method which allows one to screen putative positives from differential display analysis using only micrograms of total RNA. More importantly, we demonstrate that cDNA probes generated from amplified RNA are representative of the starting mRNA population and can be used for differential screening of mRNA species at a detectable limit of sensitivity of>/=1/40 000.
摘要
差异显示的主要障碍并非技术本身，而是差异显示后的问题，即区分假阳性和真正差异表达的mRNA。这个过程艰巨且需要大量RNA。我们提出并验证了一种方法，该方法仅使用微克级的总RNA就能从差异显示分析中筛选出假定的阳性结果。更重要的是，我们证明从扩增RNA生成的cDNA探针代表起始mRNA群体，可用于以≥1/40 000的可检测灵敏度对mRNA种类进行差异筛选。
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本文引用的文献
1
Cloning differentially expressed mRNAs.克隆差异表达的信使核糖核酸
Nat Biotechnol. 1996 Dec;14(13):1685-91. doi: 10.1038/nbt1296-1685.
2
mRNA differential display: application in the discovery of novel pharmacological targets.mRNA差异显示：在新型药理学靶点发现中的应用。
Trends Pharmacol Sci. 1996 Aug;17(8):276-9. doi: 10.1016/0165-6147(96)40001-3.
3
Differential screening of gene expression difference enriched by differential display.通过差异显示富集的基因表达差异的差异筛选。
Nucleic Acids Res. 1996 Jun 15;24(12):2454-5. doi: 10.1093/nar/24.12.2454.
4
Amplified RNA synthesized from limited quantities of heterogeneous cDNA.从有限数量的异源互补DNA合成的扩增RNA。
Proc Natl Acad Sci U S A. 1990 Mar;87(5):1663-7. doi: 10.1073/pnas.87.5.1663.
5
Arbitrarily primed PCR fingerprinting of RNA.RNA的随机引物PCR指纹分析
Nucleic Acids Res. 1992 Oct 11;20(19):4965-70. doi: 10.1093/nar/20.19.4965.
6
Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction.通过聚合酶链反应对真核生物信使核糖核酸进行差异显示
Science. 1992 Aug 14;257(5072):967-71. doi: 10.1126/science.1354393.
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