Team:Macquarie Australia/Protocols/GibsonAssembly2

From 2012.igem.org

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<center><h1>9 Gibson Assembly Reactions (2nd Attempt)</h1></center>
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<table border="3" cellpadding="4" cellspacing="0">
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<td>1.addition of 20ng gene fragments in appropriate tube</td>
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<td>1. Addition of 20ng gene fragments in appropriate tube</td>
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<td>2 μl each of Deino_A, Deino_B, Deino_C, Deino_D, Deino_E </td>
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<td>2 μL each of Deino_A, Deino_B, Deino_C, Deino_D, Deino_E </td>
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<td>2 μl each of Deino_A, Deino_B, Deino_C, Deino_D, Deino_E </td>
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<td>2 μL each of Deino_A, Deino_B, Deino_C, Deino_D, Deino_E </td>
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<td>2 μl each of Deino_A, Deino_B, Deino_C, Deino_D, Deino_E </td>
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<td>2 μL each of Deino_A, Deino_B, Deino_C, Deino_D, Deino_E </td>
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<td>2 μl each of AgroT7_A, Agro_B, Agro_C, Agro_D, Agro_E</td>
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<td>2 μL each of AgroT7_A, Agro_B, Agro_C, Agro_D, Agro_E</td>
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<td>2 μl each of AgroT7_A, Agro_B, Agro_C, Agro_D, Agro_E</td>
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<td>2 μL each of AgroT7_A, Agro_B, Agro_C, Agro_D, Agro_E</td>
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<td>2 μl each of AgroT7_A, Agro_B, Agro_C, Agro_D, Agro_E</td>
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<td>2 μL each of AgroT7_A, Agro_B, Agro_C, Agro_D, Agro_E</td>
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<td>2 μl each of Agro_A, Agro_B, Agro_C, Agro_D, Agro_E</td>
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<td>2 μL each of Agro_A, Agro_B, Agro_C, Agro_D, Agro_E</td>
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<td>2 μl each of Agro_A, Agro_B, Agro_C, Agro_D, Agro_E</td>
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<td>2 μL each of Agro_A, Agro_B, Agro_C, Agro_D, Agro_E</td>
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<td>2 μl each of Agro_A, Agro_B, Agro_C, Agro_D, Agro_E</td>
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<td>2 μL each of Agro_A, Agro_B, Agro_C, Agro_D, Agro_E</td>
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<td>2. Addition of 0.05pmol of vector</td>
<td>2. Addition of 0.05pmol of vector</td>
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<td>2.8 µl PSB-1A3</td>
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<td>2.8 μL PSB-1A3</td>
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<td>2.9 µl PSB-1K3</td>
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<td>2.9 μL PSB-1K3</td>
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<td>2.7 µl PSB-1C3</td>
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<td>2.7 μL PSB-1C3</td>
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<td>2.8 µl PSB-1A3</td>
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<td>2.8 μL PSB-1A3</td>
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<td>2.9 µl PSB-1K3</td>
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<td>2.9 μL PSB-1K3</td>
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<td>2.7 µl PSB-1C3</td>
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<td>2.7 μL PSB-1C3</td>
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<td>2.8 µl PSB-1A3</td>
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<td>2.8 μL PSB-1A3</td>
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<td>2.9 µl PSB-1K3</td>
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<td>2.9 μL PSB-1K3</td>
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<td>2.7 µl PSB-1C3</td>
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<td>2.7 μL PSB-1C3</td>
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<td>3. Addition of Master Mix (μl)</td>
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<td>3. Addition of Master Mix (μL)</td>
<td>12.8</td>
<td>12.8</td>
<td>12.9</td>
<td>12.9</td>

Latest revision as of 11:53, 20 September 2012



9 Gibson Assembly Reactions (2nd Attempt)

3A 3K 3C 4A 4K 4C 5A 5K 5C
1. Addition of 20ng gene fragments in appropriate tube 2 μL each of Deino_A, Deino_B, Deino_C, Deino_D, Deino_E 2 μL each of Deino_A, Deino_B, Deino_C, Deino_D, Deino_E 2 μL each of Deino_A, Deino_B, Deino_C, Deino_D, Deino_E 2 μL each of AgroT7_A, Agro_B, Agro_C, Agro_D, Agro_E 2 μL each of AgroT7_A, Agro_B, Agro_C, Agro_D, Agro_E 2 μL each of AgroT7_A, Agro_B, Agro_C, Agro_D, Agro_E 2 μL each of Agro_A, Agro_B, Agro_C, Agro_D, Agro_E 2 μL each of Agro_A, Agro_B, Agro_C, Agro_D, Agro_E 2 μL each of Agro_A, Agro_B, Agro_C, Agro_D, Agro_E
2. Addition of 0.05pmol of vector 2.8 μL PSB-1A3 2.9 μL PSB-1K3 2.7 μL PSB-1C3 2.8 μL PSB-1A3 2.9 μL PSB-1K3 2.7 μL PSB-1C3 2.8 μL PSB-1A3 2.9 μL PSB-1K3 2.7 μL PSB-1C3
3. Addition of Master Mix (μL) 12.8 12.9 12.7 12.8 12.9 12.7 12.8 12.9 12.7

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