iGEM 2024 — Team nais

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{% block lead %}On this page you can introduce your team members, instructors, and advisors.{% endblock %}
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* [Summary](#summary)
* [February](#section1)
* [March](#section2)
* [April](#section2-1)
* [May](#section2-2)
* [June](#section3)
* [July](#section4)
* [August](#section5)

# Summary

Establishing engineered sweet potato is a time-intensive process due to the lengthy duration required for
seedling regeneration and starch accumulation in the storage roots. A detailed account of our project’s
experimental timeline is provided below. For specific procedural steps and experimental outcomes, please
refer to the [Protocols](https://2024.igem.wiki/nais/protocol) and
[Engineering Success sections](https://2024.igem.wiki/nais/engineering).

## February

* Visit to Science and Research Center of Shanghai Chen Shan Botanical Garden
* Brainstorming

![...](https://static.igem.wiki/teams/5399/notebook/p1.jpg)

![...](https://static.igem.wiki/teams/5399/notebook/p2.jpg)

![...](https://static.igem.wiki/teams/5399/notebook/p3.jpg)

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## March

* Embryogenic calli induction of sweet potato
* sgRNA target selection
* sgRNA design
* - sgRNA oligos synthesis

![Descriptive Text](https://static.igem.wiki/teams/5399/notebook/p4.jpg)

### April

* Embryogenic calli proliferation of sweet potato

![...](https://static.igem.wiki/teams/5399/notebook/p5.jpg)

![...](https://static.igem.wiki/teams/5399/notebook/p6.jpg)

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* Construction of the backbone vector

![Descriptive Text](https://static.igem.wiki/teams/5399/notebook/p7.jpg)

Figure. a. E. coli with psgR-Cas9-IbSBEI-sgRNA;

Figure. b. PCR product of IbSBEI-sgRNA (primer F: 5’-TGTAAAACGACGGCCAGT-3’;
primer R: 5’-CTCCCGATAGGTGATACCTG-3’);

Figure. c. E. coli with psgR-Cas9-IbSBEII-sgRNA;

Figure. d. PCR product of IbSBEII-sgRNA (primer F: 5’-TGTAAAACGACGGCCAGT-3’;
primer R: 5’-TGGAGAGCTTTTGAGATTCA-3’).

![...](https://static.igem.wiki/teams/5399/notebook/p8.jpg)

![...](https://static.igem.wiki/teams/5399/notebook/p9.jpg)

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* Construction of the plant expression vector

![Descriptive Text](https://static.igem.wiki/teams/5399/notebook/p10.jpg)

Figure. Left: E. coli with pCAMBIA-1300--IbSBEI-sgRNA-Cas9-IbSBEII-sgRNA; right:
PCR product of IbSBEI-sgRNA/ IbSBEII-sgRNA (primer as former description respectively)

* Agrobacterium tumefaciens transformation

![Descriptive Text](https://static.igem.wiki/teams/5399/notebook/p11.jpg)

Figure. Left: Agrobacterium tumefaciens with
pCAMBIA-1300--IbSBEI-sgRNA-Cas9-IbSBEII-sgRNA; right: PCR products of bacterial colonies with
IbSBEI-sgRNA/ IbSBEII-sgRNA

![...](https://static.igem.wiki/teams/5399/notebook/p12.jpg)

![...](https://static.igem.wiki/teams/5399/notebook/p13.jpg)

![...](https://static.igem.wiki/teams/5399/notebook/p14.jpg)

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* Agrobacterium-mediated transformation for sweet potato

![Descriptive Text](https://static.igem.wiki/teams/5399/notebook/p15.jpg)

* Screening and selection of calli mutant

![Descriptive Text](https://static.igem.wiki/teams/5399/notebook/p16.jpg)

![Descriptive Text](https://static.igem.wiki/teams/5399/notebook/p17.jpg)

### May

* Regeneration and sub-culture of transgenic sweet potato seedlings

![Descriptive Text](https://static.igem.wiki/teams/5399/notebook/p18.jpg)

* PCR detection for regenerated seedlings

![Descriptive Text](https://static.igem.wiki/teams/5399/notebook/p19.jpg)

Figure. a. Genetically edited sweet potato tissue culture seedlings; b. PCR
product of Hyg gene (Hyg-F: Atgaaaaagcctgaactcac; Hyg-R：ctatttctttgccctcggac); c. PCR product of
Cas9 gene (Cas9-F: GACAAGAAGTACAGCATCGG; Cas9-R: AGCTGAGACAGGTCGATC);

* Transplantation of transgenic seedlings

![...](https://static.igem.wiki/teams/5399/notebook/p20.jpg)

![...](https://static.igem.wiki/teams/5399/notebook/p21.jpg)

![...](https://static.igem.wiki/teams/5399/notebook/p22.jpg)

![...](https://static.igem.wiki/teams/5399/notebook/p23.jpg)

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## June

* Literature review

## July

* Determination of internode and stem diameter

![Descriptive Text](https://static.igem.wiki/teams/5399/notebook/p24.jpg)

Figure. a. Comparison of potted seedlings between genetically edited sweet potatoes
and the WT; b. Comparison of internode length; c. Comparison of stem diameter.

## August

* Storage roots harvest
* Determination of the expression level of SBEI and SBEII in storage roots

![Descriptive Text](https://static.igem.wiki/teams/5399/notebook/p25.jpg)

Figure. Analysis of SBEI and SBEII gene expression in genetically edited sweet
potatoes.

* Extraction of total starch in storage roots

![Descriptive Text](https://static.igem.wiki/teams/5399/notebook/p26.jpg)

* Starch analysis of transgenic sweet potato storage roots

![Descriptive Text](https://static.igem.wiki/teams/5399/notebook/p27.jpg)

![Descriptive Text](https://static.igem.wiki/teams/5399/notebook/p28.jpg)

![Descriptive Text](https://static.igem.wiki/teams/5399/notebook/p29.jpg)

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* [Abstract](#abstract)
* [1. Design](#section1)
* [2. Build](#section2)
* [3. Test](#section3)
* [4. Learn](#section4)
* [5. Future](#section5)
* [References](#references)

# Abstract

Our project aims to increase the amylose content in sweet potatoes and use them as a raw material for
developing anti-glycemic foods for people with diabetes and others needing to control blood sugar
levels. We successfully knocked out the SBEI and SBEII genes, which encode starch branching enzymes, in
sweet potatoes. The gene-edited plants showed normal growth, with stem diameters nearly doubling and
internode lengths reduced to 25% of the wild type (WT), resulting in a shorter and sturdier overall
phenotype. Testing showed that knocking out the SBE genes significantly boosted amylose content, with
the best strain achieving a 104% increase.

# 1. Design

## 1.1 Gene editing chassis selection: Sweet Potato (Ipomoea batatas)

Sweet potatoes are an excellent raw material for developing anti-glycemic foods due to their high yield
per acre, strong resistance to environmental stress, and lower susceptibility to pests and diseases [1].
Compared to corn or wheat, sweet potatoes are richer in natural vitamins (such as vitamins A and C) and
minerals (such as potassium, calcium, and iron), offering superior nutritional value [2]. Moreover,
increasing the amylose content in sweet potatoes has minimal impact on their flavor, making them more
suitable for long-term daily consumption by individuals with diabetes and others needing to control
blood sugar levels [3].

![Descriptive Text](https://static.igem.wiki/teams/5399/engineer/p1.jpg)

Figure 1. Sweet Potato Concept Map

## 1.2 Knock-out SBEI and SBEII

Foods rich in resistant starch are a large group of anti-glycemic foods. How to increase the content of
resistant starch in sweet potatoes? For this we talked to Prof. Li from Yangzhou University. We learned
that the modification methods of starch synthesis pathway in crops are relatively mature. Amylose mainly
belongs to RS2 and RS3 resistant starch, which has the property that resistant starch is not easily
digested and degraded by amylase [4]. The substrate required for the biosynthesis of both amylose and
amylopectin is ADP-glucose, and there is substrate competition between them [5]. Therefore, truncation
of the amylopectin synthesis pathway enhances the metabolic flow of amylose biosynthesis. The key
enzymes for amylopectin biosynthesis are starch branching enzymes [6]. There are two homologous genes
(SBEI and SBEII) in sweet potato that can both encode starch branching enzymes, so we will knock out
both SBEI and SBEII for increasing the yield of amylose.

![Descriptive Text](https://static.igem.wiki/teams/5399/engineer/p2.jpg)

![Descriptive Text](https://static.igem.wiki/teams/5399/engineer/p3.jpg)

Figure 2. Diagram of the biosynthetic pathways of amylose and amylopectin

## 1.3 Plasmid selection

To generate the SBEI and SBEII double mutant, the CRISPR/Cas9-mediated genome-editing system was
employed to introduce mutations in the SBEI and SBEII genes in the WT background [7]. The vectors
psgR-Cas9-At and pCambia1300 are commonly used for CRISPR/Cas9-mediated genome editing. The functional
components containing IbSBEI-sgRNA-Cas9-IbSBEII-sgRNA, were cloned into the pCambia1300 vector. This
recombinant plasmid was then transformed into sweet potato to produce the SBEI and SBEII CRISPR double
mutant.

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| ![Image 1](https://static.igem.wiki/teams/5399/engineer/p4.jpg) |
| ![Image 2](https://static.igem.wiki/teams/5399/engineer/p5.jpg) |
| ![Image 3](https://static.igem.wiki/teams/5399/engineer/p6.jpg) |

Figure 3. a. plasmid map of psgR-Cas9-IbSBEI-sgRNA; b. plasmid map of
psgR-Cas9-IbSBEII-sgRNA; c. plasmid map of psgR-pAtU6-sgRNA SBEI-pAtUBQ-Cas9-tUBQ-pAtU6-sgRNA SBE II; d.
plasmid map of pCAMBIA-1300--IbSBEI-sgRNA-Cas9-IbSBEII-sgRNA; e. Schematic diagram of the final vector
pCAMBIA-1300--IbSBEI-sgRNA-Cas9-IbSBEII-sgRNA construction

# 2. Build

## 2.1 Vector construction

We designed two pairs of sgRNA sequences targeting the knockout of the SBEI and SBEII genes using online
sgRNA design tools. To prepare the sgRNAs, we first denatured the secondary structures of the
complementary oligonucleotide strands by heating them, then gradually cooled the strands to allow
annealing and formation of double-stranded sgRNAs under T4 Polynucleotide Kinase (New England Biolabs,
UK). The vector psgR-Cas9-At was linearized using BbsI (FastDigest, Thermo Fisher, Waltham, MA, USA).
The double-stranded sgRNAs were then ligated to the linearized vector using Quick T4 DNA ligase (New
England Biolabs, UK). The recombinant vectors psgR-Cas9-IbSBEI-sgRNA and psgR-Cas9-IbSBEII-sgRNA were
subsequently transformed into E. coli (Figure 4a-4d).

To enhance gene editing efficiency, we integrated the SBEI-sgRNA and SBEII-sgRNA into a single plasmid.
The pAtU6-sgRNA-SBEII module was amplified using primers containing 5'-KpnI and 3'-EcoRI sites:

AtU6-F-KpnI: 5’-GTGGTACCCATTCGGAGTTTTTGTATCTTGTTTC-3’

chim-R-EcoRI: 5’-ACGAATTCGCCATTTGTCTGCAGAATTGGC-3’

The vector psgR-Cas9-IbSBEII-sgRNA served as the template. The amplified fragment was inserted into the
KpnI and EcoRI sites of the psgR-Cas9-At vector containing the SBEI oligos to create a construct with
both customized sgRNAs and a Cas9 module. This construct (psgR-pAtU6-sgRNA
SBEI-pAtUBQ-Cas9-tUBQ-pAtU6-sgRNA SBEII) was further digested with EcoRI and HindIII and ligated into
the pCAMBIA1300 binary vector for plant transformation.

The final construct, pCAMBIA1300-IbSBEI-sgRNA-Cas9-IbSBEII-sgRNA, was chemically transformed into E.
coli. Colony PCR was performed to confirm that the recombinant bacteria contained both the SBEI-sgRNA
and SBEII-sgRNA sequences (Figure 4e, 4f).

![Descriptive Text](https://static.igem.wiki/teams/5399/engineer/p7.jpg)

![Descriptive Text](https://static.igem.wiki/teams/5399/engineer/p8.jpg)

Figure 4. a. E. coli with psgR-Cas9-IbSBEI-sgRNA; b. PCR product of IbSBEI-sgRNA
(primer F: 5’-TGTAAAACGACGGCCAGT-3’; primer R: 5’-CTCCCGATAGGTGATACCTG-3’); c. E. coli with
psgR-Cas9-IbSBEII-sgRNA; d. PCR product of IbSBEII-sgRNA (primer F: 5’-TGTAAAACGACGGCCAGT-3’; primer
R: 5’-TGGAGAGCTTTTGAGATTCA-3’); e. E. coli with pCAMBIA-1300--IbSBEI-sgRNA-Cas9-IbSBEII-sgRNA; f.
PCR product of IbSBEI-sgRNA/ IbSBEII-sgRNA (primer as former description respectively)

## 2.2 Agrobacterium-mediated infection of callus tissues

To verify that the correct plasmid, pCAMBIA1300-IbSBEI-sgRNA-Cas9-IbSBEII-sgRNA, was transformed into
Agrobacterium tumefaciens, we used PCR to identify positive clones (Figure 5a, 5b).
Agrobacterium-mediated transformation is widely employed due to its efficiency in transforming a wide
range of plant species and producing stable transgenic lines [8].

Prior to transforming sweet potatoes, we spent over a month inducing callus tissues from the sweet
potato plants (Figure 5c, 5d). After infecting the callus tissues with Agrobacterium, we conducted
selection on media containing hygromycin to isolate the successfully transformed sweet potato tissues
(Figure 5e).

![Descriptive Text](https://static.igem.wiki/teams/5399/engineer/p9.jpg)

Figure 5. a. Agrobacterium tumefaciens with
pCAMBIA-1300--IbSBEI-sgRNA-Cas9-IbSBEII-sgRNA; b. PCR products of bacterial colonies with
IbSBEI-sgRNA/ IbSBEII-sgRNA; c. Callus liquid culture; d. Callus tissues infected by Agrobacterium,
co-culture; e. Callus tissues infected by Agrobacterium were selected using hygromycin.

## 2.3 Cultivation and genotyping of Cas-SBE

After multiple rounds of selection, regeneration, and sub-culturing of transgenic sweet potato calli, we
performed genotyping of the genetically edited plants. We extracted genomic DNA from the tissue culture
seedlings and used it as a template to amplify the hygromycin resistance gene (1026 bp) and the Cas9
gene (4101 bp). We used the hyp gene and the Cas9 gene itself as positive controls and wild type (WT)
plants as negative controls. Figures 6b and 6c show that Cas-SBE-1, Cas-SBE-2, and Cas-SBE-3 are
positive for transgenic insertion.

Further, we extracted total RNA from the positive transgenic seedlings and performed reverse
transcription to convert it into cDNA. This cDNA was then used as a template for PCR. During the PCR
amplification, fluorescent dyes on the probes bind to the template DNA and emit fluorescence. The
intensity of the fluorescence signals is proportional to the amount of DNA amplified. The PCR instrument
monitors these fluorescence signals in real time to quantify the transcription levels of SBEI and SBEII
[9].

In Cas-SBE-1, the expression levels of SBEI and SBEII were reduced by 75%. In Cas-SBE-2, the
transcription levels of SBEI decreased by 78% and SBEII decreased by 86% (Figure 6d). Therefore, the
knockout of the SBE genes has been successful.

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| --- | --- |
| ![Image 1](https://static.igem.wiki/teams/5399/engineer/p10.jpg) | ![Image 2](https://static.igem.wiki/teams/5399/engineer/p11.jpg) |

Figure 6. a. Genetically edited sweet potato tissue culture seedlings; b. PCR product of
Hyg gene (Hyg-F: Atgaaaaagcctgaactcac; Hyg-R：ctatttctttgccctcggac); c. PCR product of Cas9 gene (Cas9-F:
GACAAGAAGTACAGCATCGG; Cas9-R: AGCTGAGACAGGTCGATC); d. Analysis of SBEI and SBEII gene expression in
genetically edited sweet potatoes.

## 2.4 Phenotypic analysis of Cas-SBE

To ensure that the knockout of the SBEI and SBEII genes does not adversely affect the normal growth of
sweet potatoes and thus avoid a significant reduction in yield, we measured and analyzed several
developmental traits of the sweet potato plants in comparison with the WT. Internode length is defined
as the distance between two adjacent leaves along the stem or branch, specifically the portion of the
stem between two nodes. In the transgenic plants, the internode length was reduced by 4.5-4.7 cm
compared to the WT, while the stem diameter increased by 1.4-1.8 mm, indicating that the growth rate of
Cas-SBE plants is relatively slower and they are more robust (Figures 7a-7c) [10]. From a commercial
development perspective, these factors should be considered in cost calculations.

![Descriptive Text](https://static.igem.wiki/teams/5399/engineer/p12.jpg)

Figure 7. a. Comparison of potted seedlings between genetically edited sweet potatoes
and the WT; b. Comparison of internode length; c. Comparison of stem diameter.

# 3. Test

## 3.1 Total starch content analysis

We tested the total starch content in WT and genetically edited sweet potato lines (CAS-SBE-1,
CAS-SBE-2, CAS-SBE-3) by Total Starch Content (Enzymatic Method) Assay Kit. The results indicate a
slight increase in starch content in the CAS-SBE lines compared to the WT; however, the overall
difference is not substantial. All three CAS-SBE lines remains relatively similar, with values hovering
around 450-500 mg/g dry weight (DW), while the WT shows a slightly lower content (each of the CAS-SBE
lines represents an independent biological replicate) (Figure 8). These findings suggest that knocking
out the SBE genes does not lead to a marked change in total starch accumulation in sweet potato plants.

![Descriptive Text](https://static.igem.wiki/teams/5399/engineer/p13.jpg)

Figure 8. Total starch analysis

## 3.2 Amylose content analysis

In cases where there is no significant difference in total starch content, we have measured the
percentage of amylose in both Cas-SBE and WT. Most plants naturally store starch as a mixture of amylose
and amylopectin, with amylopectin making up most of the starch granules. Starch granules composed
entirely or almost entirely of amylose do not exist in nature because excessively high levels of amylose
can adversely affect the overall health and productivity of plants [11]. However, it is feasible to
increase the amylose content to a reasonable extent. In our gene-edited Cas-SBE plants, the proportion
of amylose increased from 21% to approximately 40%, representing an overall increase of 80%-104% (Figure
9). These results indicated that the effectiveness of our SBE gene knockout.

![Descriptive Text](https://static.igem.wiki/teams/5399/engineer/p14.jpg)

Figure 9. Percent of amylose content

## 3.3 Resistant starch content analysis

Amylose mainly belongs to RS2 and RS3 resistant starch, which has the property that resistant starch is
not easily digested and degraded by amylase [4]. Therefore, based on the increase in amylose content, we
would also expect an increase in the resistant starch content in the Cas-SBE plants. We used a resistant
starch assay kit to measure the resistant starch content in the plants. Compared to the WT plants, the
resistant starch content in the Cas-SBE plants increased by 121%-133% (Figure 10). Foods with high
resistant starch content can provide anti-glycemic effects [12].

![Descriptive Text](https://static.igem.wiki/teams/5399/engineer/p15.jpg)

Figure 10. Percent of resistant starch content

# 4. Learn

We have demonstrated that knocking out the SBEI and SBEII genes in sweet potatoes significantly
increases the amylose content without causing severe impacts on plant growth and development.

# 5. Future

Moving forward, we plan to further optimize these genetically modified sweet potatoes with the aim of
achieving an amylose content of approximately 60%. First, we will focus on identifying plants with both
SBEI and SBEII genes completely knocked out in the hexaploid variety by designing more precise and
effective sgRNAs. Second, we will explore the overexpression of genes associated with amylose synthesis,
such as granule-bound starch synthase (GBSS), to enhance amylose accumulation. It’s crucial to maintain
an appropriate level of amylose, as excessive amounts (above 70%) can hinder sweet potato development
and may cause digestive issues for consumers. This advancement will facilitate the use of high-amylose
sweet potatoes in the diets of individuals with diabetes or others who need to manage blood sugar,
making it possible to produce snacks for these people.

# References

1. WANG Xin, LI Qiang, CAO QingHe, MA DaiFu. Current Status and Future Prospective of Sweet potato
   Production and Seed Industry in China[J]. Scientia Agricultura Sinica, 2021, 54(3): 483-492.
2. Zheng Y, Wang Q, Li B, Lin L, Tundis R, Loizzo MR, Zheng B, Xiao J. Characterization and Prebiotic
   Effect of the Resistant Starch from Purple Sweet Potato. Molecules. 2016 Jul 19;21(7):932.
3. Zhong Y, Tai L, Blennow A, Ding L, Herburger K, Qu J, Xin A, Guo D, Hebelstrup KH, Liu X.
   High-amylose starch: Structure, functionality and applications. Crit Rev Food Sci Nutr.
   2023;63(27):8568-8590.
4. Birt DF, Boylston T, Hendrich S, Jane JL, Hollis J, Li L, McClelland J, Moore S, Phillips GJ,
   Rowling M, Schalinske K, Scott MP, Whitley EM. Resistant starch: promise for improving human health.
   Adv Nutr. 2013 Nov 6;4(6):587-601.
5. Changsheng Li, Yongcai Huang, Ruidong Huang, Yongrui Wu, Wenqin Wang. The genetic architecture of
   amylose biosynthesis in maize kernel. Plant biotechnology journal. 2018;16(2):688-695.
6. Wang H, Wu Y, Zhang Y, Yang J, Fan W, Zhang H, Zhao S, Yuan L, Zhang P. CRISPR/Cas9-Based
   Mutagenesis of Starch Biosynthetic Genes in Sweet Potato (Ipomoea Batatas) for the Improvement of
   Starch Quality. Int J Mol Sci. 2019 Sep 23;20(19):4702.
7. Wenshan Liu, Xiaohong Zhu, Mingguang Lei, Qingyou Xia, Jose Ramon Botella, Jian-Kang Zhu, Yanfei
   Mao. A detailed procedure for CRISPR/Cas9-mediated gene editing in Arabidopsis thaliana. Science
   Bulletin. 2015, 60(15):1332-1347.
8. Zambryski, P., Joos, H., Genetello, C., Leemans, J., Montagu, M. V., & Schell, J. (1983). "Ti
   plasmid vector for the introduction of DNA into plant cells without alteration of their normal
   regeneration capacity." EMBO Journal, 2(12), 2143–2150.
9. Livak, K.J., & Schmittgen, T.D. (2001). "Analysis of relative gene expression data using real-time
   quantitative PCR and the 2−ΔΔCT method." Methods, 25(4), 402-408.
10. Davies, P. J. (2010). "Plant Hormones: Biosynthesis, Signal Transduction, Action!" Springer.
11. Richard F. Tester, John Karkalas, Xin Qi. Starch—composition, fine structure and architecture.
    Journal of Cereal Science. 2024, 39(2):151-165.
12. Wang S, De Paepe K, Van de Wiele T, Fu X, Wang S, Zhang B, Huang Q. Starch-entrapped microspheres
    enhance gut microbiome-mediated anti-obesity effects of resistant starch in high-fat diet induced
    obese C57BL/6J mice. Food Res Int. 2023 Oct;172:113215.

{% endblock %}

{% extends "layout.html" %}
{% block title %}Experiments{% endblock %}
{% block lead %}Describe the research, experiments, and protocols you used in your iGEM project.{% endblock %}
{% block page\_content %}

## What should this page contain?

---

Describe the research, experiments, and protocols you used in your iGEM project. These should be detailed enough for another team to repeat your experiments.

If you made Parts this year, please remember to put all information, characterization, and measurement data on the Part's Main Page on the [Registry](https://parts.igem.org/Main_Page).

## Inspirations

---

* [2019 Nantes](https://2019.igem.org/Team%3ANantes/Experiments)
* [2019 TU Eindhoven](https://2019.igem.org/Team%3ATU_Eindhoven/Experiments)
* [2019 Mingdao](https://2019.igem.org/Team%3AMingdao/Demonstrate)
* [2020 Amsterdam](https://2020.igem.org/Team%3AAmsterdam/Experiments)
* [2020 NCTU Formosa](https://2020.igem.org/Team%3ANCTU_Formosa/Experiments)
* [2020 USAFA](https://2020.igem.org/Team%3AUSAFA/Experiments)

{% endblock %}

{% extends "layout.html" %}
{% block title %}Results{% endblock %}
{% block lead %}You can describe the results of your project and your future plans here.{% endblock %}
{% block page\_content %}

## What should this page contain?

---

* Clearly and objectively describe the results of your work.
* Future plans for the project.
* Considerations for replicating the experiments.

## Describe what your results mean

---

* Interpretation of the results obtained during your project. Don't just show a plot/figure/graph/other, tell us what you think the data means. This is an important part of your project that the judges will look for.
* Show data, but remember **all measurement and characterization data must also be on the Part's Main Page on the [Registry](https://parts.igem.org/Main_Page).** Otherwise these data will not be in consideration for any medals or part awards!
* Consider including an analysis summary section to discuss what your results mean. Judges like to read what you think your data means, beyond all the data you have acquired during your project.

## Project Achievements

---

You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.

* A list of linked bullet points of the successful results during your project
* A list of linked bullet points of the unsuccessful results during your project and what you learned from them. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort. Furthermore, you can still impress the judges by thoughtfully discussing what went wrong and how you might change things.

## Inspirations

---

* [2019 Newcastle](https://2019.igem.org/Team%3ANewcastle/Results)
* [2019 Munich](https://2019.igem.org/Team%3AMunich/Results)
* [2019 Tec Chihuahua](https://2019.igem.org/Team%3ATec-Chihuahua/Results)
* [2020 Aalto Helsinki](https://2020.igem.org/Team%3AAalto-Helsinki/Results)
* [2020 GreatBay SCIE](https://2020.igem.org/Team%3AGreatBay_SCIE/Results)
* [2020 Queens Canada](https://2020.igem.org/Team%3AQueens_Canada/Results)

{% endblock %}