Difference between revisions of "Part:BBa K4712063"
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<partinfo>BBa_K4712063 parameters</partinfo> | <partinfo>BBa_K4712063 parameters</partinfo> | ||
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| − | + | Protocol: | |
| + | 1. For a 20μL reaction system: | ||
<table><tr><th>Reagent</th><th>Stock Concentration</th><th>Volume Added（μL）</th></tr><tr><td>Forward Primer</td><td>10μM</td><td>1</td></tr><tr><td>Reverse Primer</td><td>10μM</td><td>1</td></tr><tr><td>Rehydration Buffer (2X)</td><td></td><td>10</td></tr><tr><td>DNA Template</td><td>10nM/L</td><td>2</td></tr><tr><td>ddH2O</td><td></td><td>To 18</td></tr><tr><td>Starter (10X)</td><td></td><td>2</td></tr></table> | <table><tr><th>Reagent</th><th>Stock Concentration</th><th>Volume Added（μL）</th></tr><tr><td>Forward Primer</td><td>10μM</td><td>1</td></tr><tr><td>Reverse Primer</td><td>10μM</td><td>1</td></tr><tr><td>Rehydration Buffer (2X)</td><td></td><td>10</td></tr><tr><td>DNA Template</td><td>10nM/L</td><td>2</td></tr><tr><td>ddH2O</td><td></td><td>To 18</td></tr><tr><td>Starter (10X)</td><td></td><td>2</td></tr></table> | ||
| + | 2. Gently tap to mix several times, briefly centrifuge, repeat 3 times (mix gently to avoid vigorous vortexing). | ||
| + | 3. Incubate at 37°C for 20 minutes. | ||
| + | 4. After heating at 65°C for 10 minutes, proceed to gel electrophoresis. | ||
Revision as of 11:42, 12 October 2023
NME-R4
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 Neisseria meningitidis DNA or RNA sequences. The primers provided data for mathematical modeling for further primer design.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Protocol:
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 |
| ddH2O | To 18 | |
| Starter (10X) | 2 |
2. Gently tap to mix several times, briefly centrifuge, repeat 3 times (mix gently to avoid vigorous vortexing). 3. Incubate at 37°C for 20 minutes. 4. After heating at 65°C for 10 minutes, proceed to gel electrophoresis.