DNA Amplification

DNA probe / nucleic acid probe

● Product Name: Magigen DNA Probe / FAM fluorescence probes 100p

● Catalog#: C009S

● Size: 100 pmol (50 Test)

● DNA Probe Price: 19.9 USD

● Contents

● Advantages and Features

In the CRISPR/Cas detection system, when CRISPR/Cas protein recognizes and cleaves target double stranded DNA guided by guide RNA, its accessory cleavage activity is activated, which can efficiently cleave non-specific single stranded DNA (ssDNA) in the system.

The detection and signal amplification of DNA templates by crispr/cas can be achieved by designing ssDNA probes with Fluorophores or other markers labeled at both ends. Common detection methods include real-time fluorescence and colloidal gold chromatography.

At present, the system has been widely applied in gene editing research of microorganisms, plants, and animals, and has great potential in molecular diagnosis.

● Applicable instruments

Microplate reader, such as BioTek Synergy, or quantitative PCR instruments with FAM fluorescence channels.

● Test Steps for Fluorescence probe method*

1. Reaction substrates preparation

    The reaction substrate can be the nucleic acid amplification product of LAMP/PCR/RPA and other   amplification methods.

2. CRISPR/Cas reaction system

Note:

When using the microplate reader, set the program in advance and warm up.

Filter set: Green (Ex: 485/20, Em: 528/20), reaction temperature: 37 ℃, read the fluorescence value every 1 ~ 4 min, time: 30 min ~ 1H.

3. Result analysis

The positive or negative result of CRISPR/Cas reaction is shown in the following figure.

● Application

This nucleic acid probe is used for the development of testing products on the CRISPR technology platform.

● Storage Conditions and Shelf Life

Sealed and stored at -20 ℃,   12 months.

● Cautions

1. This DNA probe is for scientific research use only. Please read this manual carefully before use.

2. Please strictly follow the management standards of the gene amplification laboratory for experimental operations, and pay attention to preventing contamination of amplification products.

3. After the experiment, the waste generated during the testing process should be disposed of in accordance with relevant regulations.

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Tips

*FAM

Fluorescein amidites, abbreviated as FAM, are important synthetic equivalents of fluorescein dye used in oligonucleotide synthesis and molecular biology. FAM is used in the preparation of fluorescein-labeled oligonucleotide probes for the detection of the presence of the complementary nucleic acids or primers for polymerase chain reaction. Oligonucleotides labeled with fluorescein at one of the termini and with a quencher at the other can serve as molecular beacons.

FAM is the most commonly used fluorescent dye attachment for oligonucleotides and is compatible with most fluorescence detection equipment. It becomes protonated and has decreased fluorescence below pH 7; it is typically used in the pH range 7.5–8.5. FAM can be attached to 5' or 3' end of oligos.

*What is Fluorescence probe method?

Fluorescence probe method means that the photophysical properties of probe molecules change obviously before and after the specific binding between probe and analyte, so as to detect the change of fluorescence signal, realizing the detection of different molecular or ion content in the organism or the environment.