Kit for quantitatively detecting ABL mRNA level

A quantitative detection and kit technology, applied in the field of kits for quantitative detection of ABL mRNA levels, can solve the problems of unintuitive and difficult to understand the results of unknown samples, and achieve the effects of low cost, long storage period and wide coverage

Inactive Publication Date: 2010-01-13
秦亚溱 +2
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Problems solved by technology

[0003] To calculate the amount of starting template, a standard curve must be used. The standard curve is made by RQ-PCR using serially diluted standards of known starting amount. The standard can be cDNA of known quality or known copy number. The former is relatively simple to make, but the result of the

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  • Kit for quantitatively detecting ABL mRNA level

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Embodiment 1

[0023] Embodiment 1, the preparation of the kit of quantitative detection ABL mRNA level

[0024] 1. Design of primers and probes in the kit for quantitative detection of ABL mRNA levels

[0025] The composition of the real-time quantitative PCR system is as follows:

[0026] (1) Upstream primer (located in ABL exon 2):

[0027] 5'-TGGAGATAACACTCTAAAGCATAACTAAAGGT-3' (sequence 1 in the sequence listing), the final concentration is 0.3 μM;

[0028] (2) Downstream primer (located at ABL exon 3): 5'-GATGTAGTTGCTTGGGACCCA-3' (sequence 2 in the sequence listing), the final concentration is 0.3 μM;

[0029] (3) TaqMan probe (located in ABL exon 3): 5'-CCATTTTTGGTTTGGGCTTCACACCATT-3' (sequence 3 in the sequence listing), the final concentration is 0.2 μM;

[0030] (4) Master Mix for fluorescent PCR (purchased from ABI, USA).

[0031] The 3' end of the TaqMan probe in the real-time quantitative PCR system for internal reference genes is connected with a fluorescent quenching group...

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Abstract

The invention discloses a kit for quantitatively detecting ABL mRNA level. The kit comprises a standard product for manufacturing a standard curve and an internal reference gene real-time quantitative PCR system. The kit can accurately, rapidly and quantitatively detect the ABL mRNA level. In various leukaemia fusion gene detections, ABL is taken as the internal reference gene, the kit is used for performing quantitation of the internal reference gene to realize accurate, rapid and quantitative detection of various leukaemia fusion gene, further accurately and rapidly performs molecular diagnosis of leukaemia and monitoring of minimal residual diseases during treatment, and provides important molecular basis for confirmed diagnosis of clinic diseases, determination of treatment proposals, curative effect evaluation and prognosis.

Description

technical field [0001] The invention relates to a kit for quantitatively detecting the level of ABL mRNA. Background technique [0002] The real-time quantitative PCR technology (RQ-PCR) invented in recent years has realized the leap from qualitative to real quantitative PCR. Compared with ordinary PCR, RQ-PCR has the following advantages in addition to being able to quantify: (1) By adding probes to the PCR reaction system or by making a melting curve, the specificity and sensitivity are enhanced; (2) the amplification Closed tube operation during the process collects data in real time, reducing chances of contamination and false positive results; (3) no need for electrophoresis, shortening operating time; (4) evaluating sample quality by quantifying internal reference genes, reducing false negative results. The key point of the RQ-PCR technology based on TaqMan probes is that in addition to the upstream and downstream primers in the PCR reaction system, TaqMan probes are ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 秦亚溱刘艳荣主鸿鹄
Owner 秦亚溱
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