Part:BBa_K4712014

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

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE 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:

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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.

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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