PCR reagent indicator
A technology of indicator and color indicator, applied in the direction of microbial determination/inspection, biochemical equipment and methods, etc., can solve the problems of increasing detection cost and detection time, and achieve the effect of saving extraction time
Inactive Publication Date: 2015-03-25
SHANGHAI ADICON CLINICAL LAB LNC
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AI Technical Summary
Problems solved by technology
[0006] However, the abnormal pH value of the nucleic acid lysate cannot be known through a simple method before the PCR test, but can only be known after the PCR test is completed, thus increasing the test cost and test time
Method used
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Experimental program
Comparison scheme
Effect test
example 1
[0035] In TOYOBO's commercially available Taq reagent, add neutral red indicator to the PCR buffer system alone: in the color change range of 6.4-8.0, as the pH value increases, the color develops to yellow.
[0036]
example 2
[0038] Add bromocresol green to the Taq reagent commercially available from QIAGEN separately in the PCR buffer system: it is green, and can turn yellow when it is weakly acidic.
[0039]
example 3
[0041] Add phenol red to the PCR buffer system separately in the Taq reagent commercially available from TAKARA: pay attention to the color, the acid is yellow, and the alkali is red.
[0042]
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The invention relates to a PCR reagent indicator which comprises a PCR reagent and a color indicator. The color indicator represents the PH value of a lysate after DNA splitting of the PCR reagent by virtue of color. Existing fluorescent quantitative PCR reagents are colorless transparent reagents. When extraction fails, an interior label is just added to detect. The method detects by virtue of PH indication and can discover the problem of high and low PH of sample extraction which leads to PCR amplification failure. The pH indicator is added, and when the PH of a PCR reaction liquid exceeds the range, the condition can be immediately discovered by change of the color, so that the re-extraction time after failure is shortened.
Description
[0001] technical field [0002] The invention relates to a PCR buffer system, in particular to a specific pH color indicator for judging whether the pH value of the buffer system exceeds the requirement. [0003] Background technique [0004] Since the existing fluorescent quantitative PCR reagents generally use heat-resistant polymerases such as TAQ, this type of polymerase has a characteristic that it is necessary to provide a buffer system to adjust and improve the pH value that is most conducive to the activity of the polymerase. Complete PCR testing. [0005] In the existing clinical rapid detection kits, many of them also use PEG precipitation surfactant to lyse, but the pH value in the blood is abnormal or other problems cause the pH value of the final nucleic acid lysate to be high (>8.5) or low (<6.5) ), it may cause PCR detection to be inhibited, and in severe cases, it will cause PCR failure. [0006] However, the abnormal pH value of the nucleic acid ...
Claims
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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2527/119
Inventor 陈坚黎飒
Owner SHANGHAI ADICON CLINICAL LAB LNC