Team:Heidelberg/Delftibactin/DelH

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                         <h1><span style="font-size:180%;color:#FFCC00;">Del H.</span><span class="text-muted" style="font-size:120%"> This bitchy 18 kbp fragment.</span></h1>
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                         <h1><span style="font-size:180%;color:#FFCC00;">Del H.</span><span class="text-muted" style="font-size:120%"> This nasty 18 kb fragment.</span></h1>
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                         <p style="font-size:14px">Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet.</p>
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                         <p style="font-size:14px">Facing the challenge to clone 18 kbp of genomic DNA from <i>D. acidovorans</i>.</p>
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             <!--Months-->
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                                   <h1>Week 4</h1>
                                   <h1>Week 4</h1>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">The elongation of the pSB6A1-AraC-lacZ backbone turned out to be difficult. So in week 4, we performed different restriction digests both with our final backbone pSB6A1-AraC-lacZ and with the former construct pSB1C3-AraC-lacZ to verify the identity of the backbone. Besides, the amplification of the DelH fragment F1 was planned: it will be amplified in 2 subfragments - fragment F1a and fragment F1b. Each fragments has the size of 5 kb. </p>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">The elongation of the pSB6A1-AraC-lacZ backbone turned out to be difficult. So in week 4, we performed different restriction digests both with our final backbone pSB6A1-AraC-lacZ and with the former construct pSB1C3-AraC-lacZ to verify the identity of the backbone. Besides, the amplification of the DelH fragment F1 was planned: it will be amplified in 2 subfragments - fragment F1a and fragment F1b (both 5 kb in size). </p>
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                                       <h1>Week 9</h1>
                                       <h1>Week 9</h1>
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                                   <h1>Week 11</h1>
                                   <h1>Week 11</h1>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">Based on this week's experiments and in silico analysis to validate the fragments DelH F1a, F1b and F2, we discarded the restriction digest and ligation strategy for the assembly of the DelH plasmid. Instead, we chose Gibson assembly as alternative method, developed a strategy and designed primers accordingly. We are positive, to finally assemble pHM02.  
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">Based on this week's experiments and <i>in silico</i> analysis of the fragments DelH F1a, F1b and F2, we discarded the restriction digest and ligation strategy for the assembly of the DelH plasmid. Instead, we chose Gibson assembly as an alternative method, developed a strategy and designed primers accordingly.  
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                                   <h1>Week 12</h1>
                                   <h1>Week 12</h1>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">The primers arrived and we started amplifying the Gibson fragments using different approaches. Since G1 could not be PCR amplified, we decided to divide G1 into sub fragments G1a and G1b, for which we had designed and ordered primers already. The amplification of fragment G2 worked well and was produced in reasonable amounts. Alternatively, we also produced the entire DelH as one fragment G0, as well as further sub fragments DelH G1/2a, G1b/2 and G2b. Amplification of G1b/2a did not work out. Due to the failure of the restriction digest strategy, we further analyzed the backbone pSB6A1-AraC-lacZ, which we decided to discard and instead, use pSB6A1 and BBa_J04450, which is already available in the parts registry. </p>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">We started amplifying the Gibson fragments using different PCR approaches. Since we were not able to amplify G1 by PCR, we decided to divide G1 into the subfragments G1a and G1b, for which we had already designed and ordered primers. The amplification of fragment G2 worked well and yielded sufficient DNA amounts for subsequent steps. Alternatively, we also produced the entire DelH G0 as one fragment, as well as further subfragments DelH G1/2a, G1b/2 and G2b. Amplification of G1b/2a did not work out. Due to the failure of the restriction digest strategy, we further analyzed the backbone pSB6A1-AraC-lacZ, which we decided to discard. Instead, we choose to use pSB6A1 and BBa_J04450, which is already available in the parts registry. </p>
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                                   <h1>Week 13</h1>
                                   <h1>Week 13</h1>
                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">In order to realize the new strategy using the already existing backbone from the parts registry pSB6A1-lacZ-mRFP, we designed two new primers for the backbone amplification and created a map of pHM03.
                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">In order to realize the new strategy using the already existing backbone from the parts registry pSB6A1-lacZ-mRFP, we designed two new primers for the backbone amplification and created a map of pHM03.
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The plasmid for the backbone was obtained from the registry, transformed in E.coli and minipreped. The Gibson fragment of the backbone was successfully amplified, together with Gibson fragments DelH G0, G1 and G2b. </p>
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The plasmid for the backbone was obtained from the registry, transformed into <i>E.coli</i> and miniprepped. The Gibson fragment of the backbone was successfully amplified, together with Gibson fragments DelH G0, G1 and G2b. </p>
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                                   <h1>Week 14</h1>
                                   <h1>Week 14</h1>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">In week 14, we've tested numerous colonies from last week's Gibson assembly 28-07 using DelH G0 as well as G1/2a and 2b by screening-PCR. None of the analyzed colonies carried a correct plasmid. We therefore performed another Gibson assembly 01-08 and again screened numerous clones. We found few possibly correct ones that will be further analyzed next week.
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">In week 14, we screened numerous colonies from last week's Gibson assembly (28-07) for plasmids containing DelH G0, G1/2a and 2b by screening-PCR. None of the analyzed colonies carried the correct plasmid. We performed another Gibson assembly (01-08) and screened numerous clones. We found few possibly correct ones that will be further analyzed next week.
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Additionally, the new D. acidovorans strain SPH1, whose sequence is available in GenBank, was ordered. We will have to amplify all fragments from this strain again as soon as it arrives. </p>
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Additionally, the new <i>D. acidovorans</i> strain SPH1, whose sequence is available in GenBank, was ordered. We will amplify all fragments from the genome of <i>D. acidovorans</i> SPH1 as soon as it arrives. </p>
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                                   <h1>Week 15</h1>
                                   <h1>Week 15</h1>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">We further tested colonies from last weeks Gibson assemblies using DelH G0 as well as G1/2a and 2b. Unfortunately, they were all negative in the screening-PCR. Therefore, we amplified our Gibson fragments again and performed more Gibson assemblies. Yet again, there were no positive colonies in the colony-PCR. </p>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">We screened colonies from last weeks Gibson assemblies (01-08) for plasmids containing DelH G0 as well as G1/2a and 2b. None of the screening-PCRs yielded the expected DNA bands. Therefore, we amplified the Gibson fragments again and performed further Gibson assemblies. Yet again, we could not detect positive colonies by colony-PCR. </p>
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                                   <h1>Week 16</h1>
                                   <h1>Week 16</h1>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">We further characterized the DelH plasmid created using Gibson assembly. Unfortunately, none of the screened colonies was for sure positive. Selection of red colonies was not clear, and PCR screened colonies were all negative.
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">We further characterized the DelH plasmid created by Gibson assembly. None of the screened colonies yielded definit positive results. Selection of red colonies was not clear, and PCR screened colonies were all negative.
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In order to avoid high background during screening of the colonies, we decided to run two different strategies. For first approach pHM04, we will amplify the backbone without mRFP, avoiding backbone reassembly due to ribosome binding site homology. In the second strategy pHM05, we will additionally introduce a tetracycline resistance, to ensure integration of the insert.
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In order to avoid high background during screening of the colonies, we decided to run two different strategies. First, we will amplify the backbone without mRFP, avoiding backbone reassembly due to ribosome binding site homology (pHM04). In the second strategy (pHM05), we will additionally introduce a tetracycline resistance to select for the insert via antibiotic resistance.
In addition to the primers for the new strategies, we designed a new screening primer at the end of DelH. </p>
In addition to the primers for the new strategies, we designed a new screening primer at the end of DelH. </p>
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                                   <h1>Week 17</h1>
                                   <h1>Week 17</h1>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">After performing different Gibson assemblies and screening always resulted in negative clones, the possible explanation could be a religation of the backbone fragment pSB6A1 allowing E.coli to survive and express mRFP. This week, our aim is to design a new construct without mRFP, so that we can exclude the red colonies from the screening. Therefore, we are going to use a new reverse primer for the backbone including only the terminator of the mRFP, but not the mRFP. The primers for the backbone are HM11 & HM17. The construct is named pHM04 and further explained in week 16. </p>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">A possible explanation for the failed Gibson assemblies could be a religation of the backbone fragment pSB6A1 allowing <i>E.coli</i> to survive. In this case, transformed bacteria will still express mRFP. This week, our aim is to design a new construct without mRFP (pHM04), by which we will be able to exclude red colonies from the screening. For this strategy, we are going to use a new reverse primer for the backbone still including the terminator of mRFP, but omitting mRFP itself. The primers for the backbone amplification are HM11 & HM17.</p>
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                                   <h1>Week 18</h1>
                                   <h1>Week 18</h1>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">Because the constructing pHM04 did not result in any positive clone (see experiments week 17), we will follow the idea to introduce a tetracycline resistance to the ampicillin backbone as additional selection marker for successful assembly. So we have to screen only colonies, which are white (because of the exclusion of mRFP) and grow on plates containing tetracycline. Furthermore, we decided to amplify DelH in various fragments to increase Gibson assembly efficiency. Therefore, we also ordered new screening primers. The primers were designed as shown in the following table. </p>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">Since the assembly strategy for pHM04 did not yield positive clones (see experiments week 17), we will follow the idea to introduce a tetracycline resistance to the ampicillin backbone as an additional selection marker for successful assembly. With this approach, positive clones can be easily determined by their white phenotype (exclusion of mRFP) and their ability to grow on plates containing tetracycline. Furthermore, we decided to amplify DelH in various fragments to increase Gibson assembly efficiency. Therefore, we also ordered new screening primers. The primers were designed as shown in the following table. </p>
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                                   <h1>Week 19</h1>
                                   <h1>Week 19</h1>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">After a successful electroporation, we screen numerous clones by colony-PCR and test restriction digest (PvuI-HF). Positive clones are send for sequencing. The sequencing tells us if the DelH Gibson assembly actually worked. Therefore, we send the midipreped plasmids with the reverse DN07 primer or VF2, covering the start sequence of DelH and the end of the pSB6A1 backbone without mRFP (pHM04).
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">After successful electroporation, we screened numerous clones by colony-PCR and test restriction digest (PvuI-HF). Midiprepped DNA dervied from clones positive for both methods were sent for sequencing. By sequencing of the transition sequence from the end of the pSB6A1 backbone without mRFP (pHM04) to the beginning of DelH, the assembly success of the Gibson assembly can be determined (Primer: reverse DN07 primer or VF2).Sequencing results showed truncating mutation at the beginning of DelH (in the primer region) for all clones. </p>
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The screening-PCR showed different positive colonies. After performing the restriction digest, some colonies were discarded and the rest was sent in for sequencing. We did not find any positive clone without a truncating mutation at the beginning of DelH (in the primer region). </p>
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                                   <h1>Week 20</h1>
                                   <h1>Week 20</h1>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">Again, the screening-PCRs resulted in many positive colonies. After the restriction digest, many colonies were kicked out. The remaining minipreped colonies were sent in for sequencing. Yet, we did not identify any positive clone which had no mutation at the beginning of DelH (in the primer region). </p>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">As in week 19, the screening-PCRs showed colonies positive for the DelH contatining plasmid, whereas the restriction digest reveiled many of the screening results to be false positive. The remaining miniprepped colonies were sent in for sequencing. Sequencing results showed again truncating mutation at the beginning of DelH (in the primer region) for all clones. </p>
                                    
                                    
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                                   <h1>Week 21</h1>
                                   <h1>Week 21</h1>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">Frustratingly, none of the analyzed clones showed a correct sequence. Probably, DelH by itself is toxic for E.coli and thus, only aberrant clones survive. We suspect the low quality of primers as reason for high number of mutations. Therefore, we will order HPLC purified primers.
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">None of the analyzed clones showed a correct sequence, which lead as to the assumptions following assumptions. First, we suspect DelH to be toxic for <i>E.coli</i>, thus only clones carrying the mutant DelH-plasmid survive. Second, we consider the low quality of the gibson primers as a possible explanation for the high number of mutations in the assemblies. To circumwent the latter problem, we will order HPLC purified primers.
Additionally, we tried to eliminate the mutations in DelH clones I 6b and 15 by mutagenesis. Herefore, primers were ordered. </p>
Additionally, we tried to eliminate the mutations in DelH clones I 6b and 15 by mutagenesis. Herefore, primers were ordered. </p>
                                 </div>
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                                   <h1>Week 22</h1>
                                   <h1>Week 22</h1>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">So far, we failed to find a single correct DelH clone. We suspect the separate DelH module to be toxic for the E. coli. Therefore, they select for mutated plasmids. In order to reduce the selection pressure, we used E. coli BL21 DE3, known for increased expression of the lac repressor. This strategy also did not result in any correct clone. One reason might be, that the E. coli BL21 DE3 we obtained actually are BL21 DE3 pLys, which are unfortunately Chloramphenicol resistant due to additional plasmid and thus not useful for screening and amplification of DelH construct.
+
                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">So far, we failed to obtain a single correct DelH clone. We suspect the DelH module to be toxic for <i>E. coli</i> when transformed without the other parts of the Del cluster, thus DelH-transformed cells would select for mutated plasmids. In order to reduce the selection pressure, we used <i>E. coli</i> BL21 DE3, known for increased expression of the lac repressor. This strategy also did not result in any correct clone. One reason might be, that the <i>E. coli</i> BL21 DE3 strain we obtained was actually a BL21 DE3 pLys strain, which itself is already Chloramphenicol resistant, thus not useful for screening and amplification of the DelH construct coded for on a chloramphenicol vector.
-
We got new and correct E. coli BL21 DE3 as well as NEB turbo, which significantly overexpress the lac repressor. We however found their lacZ-controlled expression to be very leaky, in contrast to E. coli BL21 DE3.
+
We obtained the correct <i>E. coli</i> BL21 DE3 strain as well as NEB <i>E. coli</i> turbo cells, which significantly overexpress the lac repressor. We found the lacZ-controlled expression of the latter to be very leaky when compared to <i>E. coli</i> BL21 DE3.
-
Since the Gibson assembly of DelH using the HPLC purified primers also exclusively resulted in mutated clones, we developed two new cloning strategies to avoid expression of DelH. The first strategy uses a weak promoter and RBS, the second introduces DelH in a ccdB helper construct. Last but not least, we continued working on the mutagenesis approach to eliminate the mutation in clone C5. It was sent for sequencing... </p>
+
Since the Gibson assembly of DelH using the HPLC purified primers also exclusively resulted in mutated clones, we developed two new cloning strategies to avoid expression of DelH. The first strategy uses a weak promoter and ribosomal binding site, the second introduces DelH in a ccdB helper construct. Lastly, we continued working on the mutagenesis approach to eliminate the mutation in clone C5. It was sent for sequencing... </p>
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                                   <h1>Week 23</h1>
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                                   <h1>Week 25</h1>
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                                   <p style="font-size:10pt; text-align:justify; position:relative; margin-left:6%;">Cras justo odio, dapibus ac facilisis in, egestas eget quam. Donec id elit non mi porta gravida at eget metus. Nullam id dolor id nibh ultricies vehicula ut id elit.</p>
+
                                   This week we tried to clone DelH into a plasmid with neither promotor nor ribosome binding site (pFS_03). We managed to obtain two clones which did not have the mutations usually observed at the beginning of DelH. This proves that DelH is in fact toxic. In parallel we constructed another plasmid (pFS_04) into which the correct DelH should then be ligated by common restriction enzyme based cloning.
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                                   <p><a class="btn btn-large btn-primary" href="#">Browse gallery</a></p>
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                <h4>Methods:</h4>
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                                       </html>{{:Team:Heidelberg/Templates/DelH week12}}<html>
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                 <div class="jumbotron">
                 <div class="jumbotron">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
-
                          <div style="visibility:hidden;overflow:hidden;height:0;width:0;" id="test" >test
 
-
                          </div>
 
Line 688: Line 652:
                 <div class="jumbotron">
                 <div class="jumbotron">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
-
                          <div style="visibility:hidden;overflow:hidden;height:0;width:0;" id="test" >test
 
-
                          </div>
 
                         <div class="tab-content">
                         <div class="tab-content">
                            
                            
-
                             <div class="tab-pane" id="b14">
+
                             <div class="tab-pane active" id="b14">
                                 <p style="font-size:12pt; text-align:justify;">
                                 <p style="font-size:12pt; text-align:justify;">
                                       </html>{{:Team:Heidelberg/Templates/DelH week14}}<html>
                                       </html>{{:Team:Heidelberg/Templates/DelH week14}}<html>
Line 715: Line 677:
                 <div class="jumbotron">
                 <div class="jumbotron">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
-
                          <div style="visibility:hidden;overflow:hidden;height:0;width:0;" id="test" >test
 
-
                          </div>
 
                         <div class="tab-content">
                         <div class="tab-content">
-
                             <div class="tab-pane active" id="a15">
+
                             <div class="tab-pane active" id="b15">
-
                                <p style="font-size:12pt; text-align:justify;">
+
-
                                </p>
+
-
                            </div>
+
-
 
+
-
                            <div class="tab-pane" id="b15">
+
                                 <p style="font-size:12pt; text-align:justify;">
                                 <p style="font-size:12pt; text-align:justify;">
                                       </html>{{:Team:Heidelberg/Templates/DelH week15}}<html>
                                       </html>{{:Team:Heidelberg/Templates/DelH week15}}<html>
Line 746: Line 701:
                 <div class="jumbotron">
                 <div class="jumbotron">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
-
                          <div style="visibility:hidden;overflow:hidden;height:0;width:0;" id="test" >test
 
-
                          </div>
 
Line 778: Line 731:
                 <div class="jumbotron">
                 <div class="jumbotron">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
-
                          <div style="visibility:hidden;overflow:hidden;height:0;width:0;" id="test" >test
 
-
                          </div>
 
Line 785: Line 736:
                            
                            
-
                             <div class="tab-pane" id="b17">
+
                             <div class="tab-pane active" id="b17">
                                 <p style="font-size:12pt; text-align:justify;">
                                 <p style="font-size:12pt; text-align:justify;">
                                       </html>{{:Team:Heidelberg/Templates/DelH week17}}<html>
                                       </html>{{:Team:Heidelberg/Templates/DelH week17}}<html>
Line 806: Line 757:
                 <div class="jumbotron">
                 <div class="jumbotron">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
-
                          <div style="visibility:hidden;overflow:hidden;height:0;width:0;" id="test" >test
 
-
                          </div>
 
Line 838: Line 787:
                 <div class="jumbotron">
                 <div class="jumbotron">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
-
                          <div style="visibility:hidden;overflow:hidden;height:0;width:0;" id="test" >test
 
-
                          </div>
 
Line 870: Line 817:
                 <div class="jumbotron">
                 <div class="jumbotron">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
-
                          <div style="visibility:hidden;overflow:hidden;height:0;width:0;" id="test" >test
 
-
                          </div>
 
Line 902: Line 847:
                 <div class="jumbotron">
                 <div class="jumbotron">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
-
                          <div style="visibility:hidden;overflow:hidden;height:0;width:0;" id="test" >test
 
-
                          </div>
 
Line 934: Line 877:
                 <div class="jumbotron">
                 <div class="jumbotron">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
-
                          <div style="visibility:hidden;overflow:hidden;height:0;width:0;" id="test" >test
 
-
                          </div>
 
-
 
                         <div class="tab-content">
                         <div class="tab-content">
Line 955: Line 895:
                 </div>
                 </div>
             </div>
             </div>
 +
           
-
             <!--Week23-->
+
             <!-- Week 25 -->
             <div class="labjournal-weekly">
             <div class="labjournal-weekly">
                 <div>
                 <div>
                     <ul class="nav nav-tabs">
                     <ul class="nav nav-tabs">
-
                       <li class="active"><a href="#a23" data-toggle="tab">Overview</a></li>
+
                       <li class="active"><a href="#a25" data-toggle="tab">Overview</a></li>
-
                       <li><a href="#b23" data-toggle="tab">Lab book</a></li>
+
                       <li><a href="#b25" data-toggle="tab">Lab book</a></li>
                     </ul>
                     </ul>
                 </div>
                 </div>
                 <div class="jumbotron">
                 <div class="jumbotron">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
                     <div class="nav navbar" data-spy="scroll" data-target="#navbarExample" data-offset="0">
-
                          <div style="visibility:hidden;overflow:hidden;height:0;width:0;" id="test" >test
 
-
                          </div>
 
-
 
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                         <div class="tab-content">
-
                            <div class="tab-pane active" id="a23">
+
 
-
                                <h4>Amplification of 1 </h4>
+
                              <div class="tab-pane active" id="a25">
                                 <p style="font-size:12pt; text-align:justify;">
                                 <p style="font-size:12pt; text-align:justify;">
 +
                                  </html>{{:Team:Heidelberg/Templates/DelH_overview25}}<html>
                                 </p>
                                 </p>
                             </div>
                             </div>
-
 
+
                             <div class="tab-pane" id="b25">
-
                             <div class="tab-pane" id="b23">
+
                                 <p style="font-size:12pt; text-align:justify;">
                                 <p style="font-size:12pt; text-align:justify;">
-
                                      </html>{{:Team:Heidelberg/Templates/DelH week23}}<html>
+
                                  </html>{{:Team:Heidelberg/Templates/DelH_week25}}<html>
                                 </p>
                                 </p>
                             </div>
                             </div>
 +
                              
                              
                         </div>
                         </div>
                     </div>
                     </div>
                 </div>
                 </div>
 +
            </div>
             </div>
             </div>
Line 992: Line 932:
</div>
</div>
</html>
</html>
 +
{{:Team:Heidelberg/Templates/Footer-Nav}}
{{:Team:Heidelberg/Templates/Footer-DelH}}
{{:Team:Heidelberg/Templates/Footer-DelH}}

Latest revision as of 03:33, 29 October 2013

Del H. This nasty 18 kb fragment.

Facing the challenge to clone 18 kbp of genomic DNA from D. acidovorans.

Heidelberg ga delf.png

Thanks to