The invention relates to a reagent kit and a method of detecting KRAS gene mutation in free DNA of plasma. The reagent kit comprises a specific primer pair and a high-temperature-resistant restriction endonuclease, wherein the specific primer comprises a sense primer with a sequence shown as SEQ ID NO. 1, and an antisense primer with a sequence shown as SEQ ID NO. 2; the high-temperature-resistant restriction endonuclease is a Bsr I endonuclease. With the adoption of the reagent kit and the detection method for detecting the KRAS gene mutation, the mutation detection sensitivity can be improved; the specimen consumption is reduced; clinical utilization and popularization are facilitated.

Reagent kit and method of detecting KRAS gene mutation in free DNA of plasma

Name: a detection method of gene hot-spot mutation. Abstract: the detection method provided by the invention, by conducting restriction enzyme digestion on a wild sequence in vitro and by conducting treatment, which is not good for a ligation reaction, at the end of an incision, sub-clone an in-vitro amplification fragment, and then implements sequencing analysis on a colony formed by cloning. The detection method provided by the invention is conducive to the formation of the colony from mutant genes, while the capacity of a wild gene fragment to form a colony declines due to the restriction enzyme reaction and the ligation inhibiting treatment. The detection method provided by the invention has an application value in the aspect of analyzing gene mutation of a biological specimen which is low in mutant gene content.

Detection method of gene hot-spot mutation

The invention discloses a fragment for selectively enhancing a mutant nucleic acid by using a report gene, thereby selectively suppressing amplification of a wild gene fragment containing an allele in a nucleic acid sample and not suppressing amplification of a mutant gene fragment containing the allele in the nucleic acid sample.

Fragment for selectively enhancing mutant nucleic acid by using report gene and method of fragment

The invention relates to a primer pair for detecting KRAS hot-spot mutation by utilizing blue-white screening. The technical scheme lies in that the principle of in-vitro adjustment of the reading frame of genes is adopted, and the blue-white screening for subcloning is used for detecting hot-spot mutation of tumor mutant KRAS genes at the amino acid codons 12 and 13; the invention discloses the primer pair for detecting KRAS hot-spot mutation by utilizing blue-white screening. According to the primer pair provided by the invention, selective amplification and reading frame adjustment dual functions are realized, the KRAS hot-spot mutation can be converted into a termination codon in beta galactosidase fusion genes, therefore the screening effect that wild type gene segments form blue escherichia coli bacterial colonies after subcloning while mutant gene segments form white bacterial colonies after subcloning is achieved; besides, the primer pair has an application value in KRAS hot-spot mutation analysis.

Primer pair for detecting KRAS hot-spot mutation by utilizing blue-white screening

Handle everyday research tasks with reliable, citation-backed results

Your personal Research Agent to handle research tasks with citation-backed results

Popular Tasks used by Researchers

How can I help with your research?

Meet SciSpace

Get more enhanced response by uploading the PDFs you want me to reference.

No relevant PDFs in your library

SciSpace is the AI research assistant for academics. Run systematic literature reviews on 280M+ papers, and write papers with cited sources. Trusted by 1M+ students, PhDs & researchers.

SciSpace | AI for Research

Analyze PDFs

Code & Manuscripts

Funding & Grants

Literature & Patents

Medical & Clinical Data

Systematic Review

Visualize & Present

Web & Data

Build a Google Scholar-like website for your research.

Build a website

Create charts and images for your research

Create a Chart

Write a paper for submission to a journal

Draft a manuscript

Patent Search

Design eye-catching scientific posters in minutes.

Scientific Poster Generation

Systematic Literature Review

One task is running at the moment. Your messages will be shown right after.

Drag and drop or click here to browse

Loved by <highlight>1 million+</highlight> researchers

Extract a list of specific topics and their sources from unstructured text

Topics

Compare and analyze relevant papers that matches with your search

Papers

Get insights from PDFs and bookmarked papers from your library

My library

Recent searches

Try searching for:

Catch AI-generated content in scholarly and non-scholarly content

{ai} Detector

Ai Writer

Get PDF Summaries, highlighted text explanations 

Chat with PDF

Effortlessly create in-text citations and bibliographies in APA and 2,500 other formats

Citation generator

Get explanations, summaries, and answers on academic papers

Ease up your research workflow with {scispace}'s cohort of exciting AI tools

Elevate your academic writing skills and convey your ideas the way you want

Paraphraser

Explore our range of reading and writing tools

Your file is being prepared and should be ready in a few minutes. If it's a large file, it might take a bit longer. You can close this window, and we'll email you the file when it's done.

You have reached a maximum limit of <strong>{limit}</strong> columns in the table. Remove at least <strong>1</strong> column to add or create another one.

Li Wen

Author Tools

Chat about Author

Papers

Primer pair for detecting KRAS hot-spot mutation by utilizing blue-white screening