Primer group and kit for simultaneously amplifying 44 Y-STR gene loci of human and application thereof
A Y-STR, locus technology, applied in the field of molecular genetics, can solve the problems of inconvenience and time-consuming, and achieve the effects of high individual identification, high non-father exclusion rate, and strong amplification specificity
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Embodiment 1
[0125] Direct multiplex amplification was performed on two hairs from suspected father and son to detect 44 loci. The amplification method adopts the method of direct amplification: take a section of hair with hair follicles and put it directly into the amplification system so that the hair follicles are completely immersed in the liquid. The amplification procedure adopts the standard amplification procedure of the present invention, and the amplification instrument adopts ABI Proflex PCR instrument, genetic analyzer adopts 3500xl genetic analyzer, analysis software adopts GeneMapper-ID-X analysis software. The operation steps of this embodiment are as follows:
[0126] ①Use scissors to cut the hair short at 0.5cm above the hair follicle, and put the short hair with the hair follicle into a 200μl PCR tube.
[0127] ② According to the standard reaction system of the present invention, a 10 μl amplification system was prepared: 2 μl primer mixture, 4 μl reaction premix, 4 μl d...
Embodiment 2
[0136] The invention is mainly used for the construction of DNA database and identification.
[0137] The detailed part of the operation is as follows:
[0138] ① Known personnel samples are usually blood cards, which can be directly amplified. Use a puncher with a diameter of 1mm to directly punch a blood piece with a diameter of 1mm on the dry blood card and put it into a 200μl PCR tube.
[0139] ②Unknown persons are usually the samples collected at the scene of the case, and the samples are complicated. Usually, the method of extracting DNA first and then amplifying the data is used for detection and analysis. DNA was extracted by Chelex-100 and magnetic bead method. (For the extraction method, refer to Chapter 4 DNA Extraction of "Forensic DNA Analysis" by Zheng Xiufen).
[0140] ③ For the preparation and amplification of the reaction system, refer to the standard system of the present invention to prepare a 10 μl reaction system, see Table 7 below. PCR amplification w...
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