Team:Hawaii/Biobricks extraction & transformation competency test

From 2008.igem.org

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== Results ==
== Results ==
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<table style="text-align: left; width: 474px;" border="1"
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style="width: 121px; text-align: center; font-weight: bold;">Type
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of Cell</td>
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      <td style="width: 97px; text-align: center; font-weight: bold;">Plasmid</td>
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      <td style="width: 98px; text-align: center; font-weight: bold;">Dilution</td>
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style="text-align: center; width: 124px; font-weight: bold;">Colonies</td>
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style="width: 121px; text-align: center;">Strataclone&nbsp;
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Compentent Cells </td>
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      <td colspan="1" rowspan="2"
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style="width: 97px; text-align: center;">pUC18</td>
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      <td style="width: 98px; text-align: center;">1:1</td>
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      <td style="width: 124px; text-align: center;">Too
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many to Count</td>
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      <td style="width: 98px; text-align: center;">1:10</td>
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      <td style="width: 124px; text-align: center;">232 x 4</td>
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style="width: 97px; text-align: center;">BBa_B0034</td>
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      <td style="width: 98px; text-align: center;">1:1</td>
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      <td style="width: 124px; text-align: center;">20</td>
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      <td style="width: 98px; text-align: center;">1:10</td>
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      <td style="width: 124px; text-align: center;">1</td>
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      <td colspan="1" rowspan="4"
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style="width: 121px; text-align: center;">DH5</td>
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      <td colspan="1" rowspan="2"
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style="width: 97px; text-align: center;">pUC18</td>
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      <td style="width: 98px; text-align: center;">1:1</td>
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      <td style="width: 124px; text-align: center;">Too
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many to Count</td>
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      <td style="text-align: center;">1:10</td>
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      <td style="text-align: center;">43</td>
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      <td style="text-align: center;" colspan="1"
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rowspan="2">BBa_B0034</td>
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      <td style="text-align: center;">1:1</td>
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      <td style="text-align: center;">1</td>
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    </tr>
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    <tr>
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      <td style="text-align: center;">1:10</td>
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      <td style="text-align: center;">1</td>
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<br>
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[[Image:IMG 1672.JPG|thumb|300px|right|<please add description of what we are seeing here]]
[[Image:IMG 1672.JPG|thumb|300px|right|<please add description of what we are seeing here]]
''<colony count TBD on 6/11/08>''
''<colony count TBD on 6/11/08>''
==Discussion==
==Discussion==

Revision as of 22:21, 13 June 2008

Contents

Question

  1. Is the small amount of transformants we got on our "Initial BioBrick Plasmid Extraction From Filter Paper" experiment due to the low concentration of DNA from the filter paper or the low competency of our cells?
  2. How competent are our DH5α cells compared to commercial quality competent cells?

Protocol

The same protocol as "Initial BioBrick Plasmid Extraction From Filter Paper" was used.

  • Filter paper was smashed up really well in 7 μl TE.
  • Competency of our DH5α cells was compared to StrataCloneTM competent cells.
  • Concentration of DNA on filter paper was estimated by transforming with pUC18 (.1 ng/μg) and comparing colony counts
  • Cells were shaken at 200rpm.
  • 1/1 and 1/10 dilutions were plated.

Results

Type of Cell Plasmid Dilution Colonies
Strataclone  Compentent Cells pUC18 1:1 Too many to Count
1:10 232 x 4
BBa_B0034 1:1 20
1:10 1
DH5 pUC18 1:1 Too many to Count
1:10 43
BBa_B0034 1:1 1
1:10 1


<please add description of what we are seeing here

<colony count TBD on 6/11/08>

Discussion

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