This kit utilizes rapid nucleic acid amplification technology at room temperature or a constant temperature (typically 39ºC–42ºC). Reverse transcriptase uses specific primers and RNA templates to synthesize cDNA strands. The recombinase and primers, aided by auxiliary proteins and single-strand binding proteins, form a complex that searches for and binds to the target homology region. This results in the formation of a D-loop, initiating strand exchange. Once recombinase dissociates, polymerase binds to the 3' end of the primer and starts chain extension. With the action of the nfo enzyme, a specific molecular probe, designed according to the template, is added, and colloidal gold technology (sandwich method) is used to detect the final result.
【Features】
High sensitivity, strong specificity, and short reaction time (only 15 minutes), and the reaction components are in dry powder state, which is easy to operate and easy to store.
This reagent has low equipment requirements, and the reaction operation can be carried out in metal baths, water baths, etc., and there is no need to purchase expensive exclusive equipment such as PCR amplifiers.
Strong Specificity
This kit offers excellent performance for precise nucleic acid amplification.
Fast Reaction
The reaction completes in only 15 minutes.
Convenient
Components are in dry powder form, making them easy to store and handle.
Simple Required
Suitable for use with basic equipment such as metal or water baths, without the need for expensive PCR amplifiers.
【Primer design】
It is recommended to use primers with a length of 30-35 bp.Too short primers will affect the amplification speed and detection sensitivity;the 5' end of the reverse primer is labeled with a modification group (commonly used biotin). Primers are designed to avoid the formation of secondary structures that affect amplification;the length of the amplicon is recommended to be 150-500 bp.
【Fluorescent Probe Design】
Design a probe sequence of 46–52 nt, complementary to the target fragment, located between the upstream and reverse primers.
- An antigen tag (typically FAM) is labeled at the 5' end.
- A dSpacer (tetrahydrofuran, THF) is placed in the middle of the probe (between the 5' and 3' ends), acting as the recognition site for nfo enzyme.
- The 3' end is labeled with a modification group, such as an amine group, phosphate group, or C3-Spacer.
【Kit Composition】
|
Composition |
Content |
|
AD buffer |
1.6mL×1Tube |
|
B buffer |
150μL×1Tube |
|
Reagent |
48 T |
|
Guide Manual |
1 Copy |
Note: Considering the problem of nucleic acid degradation, RNA series products do not provide positive control templates and primers.
【Packing Specifications】
- Specifications：48 T/pack
【Kit storage】
- Storage conditions: Store at -20℃ before opening the package.
- Shelf Life: 14 months.
【Note】
- Self-designed primers/probes must be added for MIRA project development.
- The kit is available in two types - DNA and RNA. The RNA type includes reverse transcriptase, allowing amplification with RNA, dsDNA, and ssDNA as templates. The DNA type can only amplify with dsDNA as the template.
- Available in three types - Basic, Fluorescent, and Test Strip. Choose the appropriate kit based on your needs and use the corresponding primers/probes.
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