多重连接依赖探针扩增
生物
寡核苷酸
分子生物学
连接酶连锁反应
核酸
多路复用
底漆(化妆品)
分子信标
滚动圆复制
多重位移放大
聚合酶链反应
DNA
核糖核酸
计算生物学
多重聚合酶链反应
遗传学
锁核酸
分子探针
结扎
杂交探针
核酸热力学
基因
外显子
化学
DNA提取
有机化学
作者
Jan P. Schouten,Cathal J. McElgunn,Raymond Waaijer,Danny Zwijnenburg,Filip Diepvens,Gerard Pals
摘要
We describe a new method for relative quantification of 40 different DNA sequences in an easy to perform reaction requiring only 20 ng of human DNA. Applications shown of this multiplex ligation-dependent probe amplification (MLPA) technique include the detection of exon deletions and duplications in the human BRCA1, MSH2 and MLH1 genes, detection of trisomies such as Down's syndrome, characterisation of chromosomal aberrations in cell lines and tumour samples and SNP/mutation detection. Relative quantification of mRNAs by MLPA will be described elsewhere. In MLPA, not sample nucleic acids but probes added to the samples are amplified and quantified. Amplification of probes by PCR depends on the presence of probe target sequences in the sample. Each probe consists of two oligonucleotides, one synthetic and one M13 derived, that hybridise to adjacent sites of the target sequence. Such hybridised probe oligonucleotides are ligated, permitting subsequent amplification. All ligated probes have identical end sequences, permitting simultaneous PCR amplification using only one primer pair. Each probe gives rise to an amplification product of unique size between 130 and 480 bp. Probe target sequences are small (50-70 nt). The prerequisite of a ligation reaction provides the opportunity to discriminate single nucleotide differences.
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