Difference between revisions of "Team:NPU-China/Demonstrate"

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Aim to Demonstrate MitoCRAFT
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</h4>
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</div>
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<h5>
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After a year of hard work, we have been advancing the cutting edge of MitoCRAFT. <br>
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With the aid of Prof. Jiang Huifeng and his research group, we completed the entire design work of the MitoCRAFT
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genome based on the wild-type S. cerevisiae mitochondrial genome.
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</h5>
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<div class="center" style="padding-top:10px;">
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<img src="https://static.igem.org/mediawiki/2018/9/9c/T--NPU-China--model_6.png" class="responsive-img" style="width:80%">
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<h6>Fig. 1 Complete genome structure of MitoCRAFT</h6>
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</div>
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<h5>
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In particular, what we crave is not a simple mitochondrial genome, instead, we yearn to construct a standard-mode
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mitochondrial chassis that is widely used like S. cerevisiae for all sorts of functional verification and expansion
 +
related to mitochondria. In other word, we desire a powerful bio toolbox.
 +
Upon finishing the design of MitoCRAFT, we immediately started the work of de novo synthesis of the MitoCRAFT genome
 +
with corresponding primers.<br>
 +
In the first place, a series of condition optimizations have enabled us to successfully complete the de novo
 +
synthesis of 22 primary fragments. The results are as follows:
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</h5>
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<div class="center" style="padding-top:10px">
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<img src="https://static.igem.org/mediawiki/2018/8/85/T--NPU-China--jiaotu3.1.png" class="responsive-img" style="width:80%">
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<h6>Fig. 2-1 Gel electrophoresis of first-stage fragments.
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M is NormalRun™Prestained 250bp-II DNA ladder(GENERAY BIOTECH);
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L1:2778bp; L2:2265bp; L3:2883bp; L4:1759; L5:2795bp.
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</h6>
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</div>
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<div class="center" style="padding-top:10px">
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<img src="https://static.igem.org/mediawiki/2018/a/ac/T--NPU-China--jiaotu3.2.png" class="responsive-img" style="width:80%">
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<h6>Fig. 2-2 Gel electrophoresis of first-stage fragments.
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M is NormalRun™Prestained 250bp-II DNA ladder(GENERAY BIOTECH);
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L6:2288bp; L7:1142bp; L8:1368bp; L9:1453bp;L10:1376bp; L11:1904bp.
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</h6>
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</div>
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<div class="center" style="padding-top:10px">
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<img src="https://static.igem.org/mediawiki/2018/2/20/T--NPU-China--jiaotu3.3.png" class="responsive-img" style="width:80%">
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<h6>Fig. 2-3: Gel electrophoresis of first-stage fragments.
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M is NormalRun™Prestained 250bp-II DNA ladder(GENERAY BIOTECH);
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L12:2377bp; L13:1376bp; L14:1053bp; L15:1872bp; L16:1579bp; L17:1763bp.
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</h6>
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</div>
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<div class="center" style="padding-top:10px">
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<img src="https://static.igem.org/mediawiki/2018/0/0b/T--NPU-China--jiaotu4.4.png" class="responsive-img" style="width:80%">
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<h6>Fig. 2-4 Gel electrophoresis of first-stage fragments.
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M is NormalRun™Prestained 250bp-II DNA ladder(GENERAY BIOTECH);
 +
L18:2370bp; L19:3306bp; L20:2007bp; L21:525bp; L22(GFP):870bp.
 +
</h6>
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</div>
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<h5 style="padding-top:30px">
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Afterwards, the introduction of PTRCCS system, in the light of the current experimental results, initially proved
 +
the verification of the completion of the synthesis of the entire MitoCRAFT genome:
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</h5>
 +
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<h5 style="padding-top:30px">
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<b>[Results analysis]</b>
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</h5>
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<!--这里zbz说还没有写好-->
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<h5 style="padding-top:30px">
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In view of our current experimental results, we still have a lot of work to do to ultimately implement the actual
 +
application of MitoCRAFT. The difficulty of synthesizing MitoCRAFT genomes has turned out far beyond our
 +
expectation. Our team members have been conducting PCR since they entered the lab until now, but hopefully, we are
 +
really glad that our MitoCRAFT genome is almost ready to be a triumph. We also look forward to continuing to
 +
explore
 +
this cutting-edge subject after Giant Jamboree. Our specific work plans for the future have been presented on the
 +
Future Work page. We sincerely welcome all iGEMers to contact us, sharing with us your unique experience and
 +
jointly
 +
creating a bright future for MitoCRAFT and synthetic biology.
 +
</h5>
 +
 +
 +
 +
 +
 +
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{{NPU-China/down}}
 
{{NPU-China/down}}

Revision as of 01:55, 18 October 2018

menu

Aim to Demonstrate MitoCRAFT

After a year of hard work, we have been advancing the cutting edge of MitoCRAFT.
With the aid of Prof. Jiang Huifeng and his research group, we completed the entire design work of the MitoCRAFT genome based on the wild-type S. cerevisiae mitochondrial genome.
Fig. 1 Complete genome structure of MitoCRAFT
In particular, what we crave is not a simple mitochondrial genome, instead, we yearn to construct a standard-mode mitochondrial chassis that is widely used like S. cerevisiae for all sorts of functional verification and expansion related to mitochondria. In other word, we desire a powerful bio toolbox. Upon finishing the design of MitoCRAFT, we immediately started the work of de novo synthesis of the MitoCRAFT genome with corresponding primers.
In the first place, a series of condition optimizations have enabled us to successfully complete the de novo synthesis of 22 primary fragments. The results are as follows:
Fig. 2-1 Gel electrophoresis of first-stage fragments. M is NormalRun™Prestained 250bp-II DNA ladder(GENERAY BIOTECH); L1:2778bp; L2:2265bp; L3:2883bp; L4:1759; L5:2795bp.
Fig. 2-2 Gel electrophoresis of first-stage fragments. M is NormalRun™Prestained 250bp-II DNA ladder(GENERAY BIOTECH); L6:2288bp; L7:1142bp; L8:1368bp; L9:1453bp;L10:1376bp; L11:1904bp.
Fig. 2-3: Gel electrophoresis of first-stage fragments. M is NormalRun™Prestained 250bp-II DNA ladder(GENERAY BIOTECH); L12:2377bp; L13:1376bp; L14:1053bp; L15:1872bp; L16:1579bp; L17:1763bp.
Fig. 2-4 Gel electrophoresis of first-stage fragments. M is NormalRun™Prestained 250bp-II DNA ladder(GENERAY BIOTECH); L18:2370bp; L19:3306bp; L20:2007bp; L21:525bp; L22(GFP):870bp.
Afterwards, the introduction of PTRCCS system, in the light of the current experimental results, initially proved the verification of the completion of the synthesis of the entire MitoCRAFT genome:
[Results analysis]
In view of our current experimental results, we still have a lot of work to do to ultimately implement the actual application of MitoCRAFT. The difficulty of synthesizing MitoCRAFT genomes has turned out far beyond our expectation. Our team members have been conducting PCR since they entered the lab until now, but hopefully, we are really glad that our MitoCRAFT genome is almost ready to be a triumph. We also look forward to continuing to explore this cutting-edge subject after Giant Jamboree. Our specific work plans for the future have been presented on the Future Work page. We sincerely welcome all iGEMers to contact us, sharing with us your unique experience and jointly creating a bright future for MitoCRAFT and synthetic biology.