Difference between revisions of "Part:BBa K4712014"
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Revision as of 11:49, 12 October 2023
H1N1-F8
The primers were designed using NCBI BLAST and SanpGene to achieve efficient and specific amplification. This primer in RPA serves as the initial binding point for the amplification process, ensuring the specificity of the reaction by targeting the desired Influenza A virus(2009H1N1) DNA or RNA sequences. The primers provided data for mathematical modeling for further primer design.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Apparatus:
Thermal Cycler, Centrifuge, Fluorescence Quantitative PCR Instrument (Ya Rui), 2mL reaction tube, Pipette and Pipette Tip
Materials:
1. DNA Isothermal Amplification Kit (EZ-Life Biotechnology)
2. ddH2O
3. Thermostatic amplification specific primers:
INFB-F1、 INFB-R1 INFB-F2、 INFB-R2 INFB-F3、 INFB-R3 INFB-F4、 INFB-R4 INFB-F5、 INFB-R5 H1N1-F6、H1N1-R6 H1N1-F7、H1N1-R7 H1N1-F8、H1N1-R8
4. DL500 marker 5. Test sample: DNA Template(pUC57-M1)Concentration:1000cps/μL Storage: -20°C Methods: 1. For a 20μL reaction system:
Reagent | Stock Concentration | Volume Added(μL) |
---|---|---|
Forward Primer | 10μM | 1 |
Reverse Primer | 10μM | 1 |
Rehydration Buffer (2X) | 10 | |
DNA Template | 10nM/L | 2 |
H2O | To 18 | |
Starter (10X) | 2 |
2. Gently tap multiple times to mix the ingredient, and then centrifuge briefly (mix gently, avoiding vigorous shaking or vortexing). 3. Incubate at 37 ℃ for 20 minutes. 4. After heating at 65 ℃ for 10 minutes, the adhesive will run off.
Electrophoresis swimlane | Content |
---|---|
1 | DL500 maker |
2 | INFB-F1、 INFB-R1 |
3 | INFB-F2、 INFB-R2 |
4 | INFB-F3、 INFB-R3 |
5 | INFB-F4、 INFB-R4 |
6 | INFB-F6、 INFB-R6 |
7 | INFB-F5、 INFB-R5 |
8 | H1N1-F6、H1N1-R6 |
9 | H1N1-F7、H1N1-R7 |
10 | H1N1-F8、H1N1-R8 |