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Nucleic acid probe, composition containing same and application of nucleic acid probe
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A nucleic acid probe and composition technology, applied in the field of molecular biology, can solve problems such as unsatisfactory signal specificity, and achieve the effects of high specificity and high signal intensity
Inactive Publication Date: 2013-06-12
BEIJING GP MEDICAL TECH
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Currently there are relevant probes for detecting chromosome 8 (Heinz-Ulrich G. Weier, Hans-Dieter Kleine, and Joe W. Gray. Labeling of the centromeric region on human chromosome 8 by in situ hybridization[J]. Hum Genet (1991) 87: 489-494.), but the probe actually contains a mixture of multiple sequences, and the signal specificity is not ideal
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Embodiment 1
[0046] Embodiment 1: the preparation of nucleic acid probe sample 1 of the present invention
[0047] Reagents and materials:
[0048] TaKaRa Taq TM Hot Start Version, DNase I, and DNApolymerase I were purchased from Dalian TaKaRa Company; FITC-dUTP was purchased from Roche; Human Cot-1 DNA was purchased from Invitrogen; QIAprep Spin Miniprep Kit was purchased from QIAGEN.
[0049] experimental method:
[0050] 1. Acquisition of probe sequences
[0051] 1) DOP-PCR amplification of chromosome 8 DNA
[0052] Using normal human chromosome 8 DNA (microdissection) as a template, DOP-PCR amplifies chromosome 8 DNA, and the sequences of the degenerate primers used are as follows:
[0053] 5-CCGACTCGAGNNNNNNATGTGG-3 (SEQ ID NO: 2) (synthesized by TaKaRa Company), wherein N represents four bases randomly selected from ATCG.
[0149] Prepare the probe mixture according to the following ratio (take 10 people as an example) to obtain the detection agent 1, which contains the nucleic acid probe sample 1 prepared in Example 1.
[0150] Detecting agent 1 each component Volume of each component
[0156] The concentration of the nucleic acid probe sample 1 is not particularly limited, and is, for example, 0.1 μg / μl.
[0157] Although the specific numerical values for the volume of each component are given above according to 10 servings, those skilled in the art can understand that the volume of each component mainly meets the above ratio. It is also possible to prepare detection reagents for one person or multiple people.
[0158] In order t...
Embodiment 3
[0159] Embodiment 3: Preparation of control probe 1 and control probe 2
[0160] Control probe 1 was prepared in a similar manner to Example 1, except that in step 2) of obtaining the probe sequence, a single copy (171 bp) of α-satellite DNA was selected as the probe sequence.
[0161] Control probe 2 (actually a mixture of various sequences) was prepared according to the method in the literature in the background art.
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Abstract
The invention belongs to the technical field of molecular biology, relating to a nucleic acid probe, a composition containing the same and application of the nucleic acid probe. Particularly, the invention relates to a nucleic acid probe for detecting the number of chromosome No.8, and the nucleotide sequence of the nucleic acid probe can be SEQ ID NO:1 or a complementary sequence of the SEQ ID NO:1 or a variant with sequence sameness with the SEQ ID NO:1. The invention further relates to a detection agent for detecting the number of the chromosome No.8, a kit for detecting the number of the chromosome No.8, an in-situ hybridization method, a method for detecting the number of the chromosome No.8, application of the nucleic acid probe to preparation of a reagent for detecting the number of the chromosome No.8, and application of the nucleic acid probe to preparation of reagents or auxiliary reagents for detecting diseases related to abnormality of the number of the chromosome No.8. The nucleic acid probe disclosed by the invention has high signal strength and high specificity.
Description
technical field [0001] The invention belongs to the field of molecular biology and relates to a nucleic acid probe, in particular to a nucleic acid probe for detecting the number of chromosome 8. The present invention also relates to a composition containing the nucleic acid probe, a detection agent for detecting the number of chromosome 8, a kit for detecting the number of chromosome 8, an in situ hybridization method, and a method for detecting the number of chromosome 8 The method, the use of the nucleic acid probe in the preparation of a reagent for detecting the number of chromosome 8, and the use of the nucleic acid probe in the preparation of a reagent or auxiliary reagent for detecting diseases related to the abnormal number of chromosome 8. Background technique [0002] Studies have found that many diseases are accompanied by abnormal numbers of chromosome 8, such as myelodysplastic syndromes (myelodysplastic syndromes, MDS) and cancers such as acute leukemia (acute...
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