iGEM 2017 — Team Hong_Kong_UCCKE

[![](https://static.igem.org/mediawiki/2017/8/8b/HQ_page_logo.jpg)](https://2017.igem.org)

# Team:Hong Kong UCCKE/Notebook

Toggle navigation     [UCCKE 2017](https://2017.igem.org/Team%3AHong_Kong_UCCKE)

* About
  + [Team](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Team)
  + [Collaborations](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Collaborations)
  + [Contact](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Contact)
* Project
  + [Description](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Description)
  + [Design](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Design)
  + [Experiments](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Experiments)
  + [Results](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Results)
  + [Contribution](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Contribution)
  + [Notebook](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Notebook)
  + [Protocols](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Protocols)
  + [Attributions](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Attributions)
* Parts
  + [Parts](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Parts)
  + [Basic Parts](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Basic_Part)
  + [Composite Parts](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Composite_Part)
* [Safety](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Safety)
* Human Practices
  + [Silver HP](https://2017.igem.org/Team%3AHong_Kong_UCCKE/HP/Silver)
  + [Public Engagement](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Engagement)
* [Judging Form](https://igem.org/2017_Judging_Form?team=Hong_Kong_UCCKE)

## Notebook

Click on section titles to see the contents.

### Week 1 - 8 (1/12/2016 - 21/1/2017)

Oops... we haven't started preparing... consider start searching from Week 9?

### Week 9 (22-28/1/2017)

2nd iGEM Meeting
Location: Lab
Date: 23/1/2017
Time: 16:15-17:45
Attendance: All helpers
In today’s meeting, we taught new members some basic knowledge about genetic engineering,
including promoter, terminator, RBS etc.. All members were assigned to design at least one part
as homework and contribution to the team

### Week 10 (29/1/2017-4/2/2017)

Uh oh... we were lazy this week... wanna see what's happening next week?

### Week 11 (5-11/2/2017)

3rd iGEM Meeting
Location: Lab
Date: 6/2/2017
Time: 16:15-18:00
Attendance: Most of the members
In today’s meeting, we taught new members how to do gel electrophoresis.
They were taught the purpose of doing gel electrophoresis and they gave a try on doing it.
Core members in the team had meeting allocating manpower and set a solid schedule.

### Week 12 (12-18/1/2017)

4th iGEM Meeting
Location: Lab
Date: 13/2/2017
Time: 16:15-17:50
Attendance: Most of the members
We had a regular meeting today.

### Week 13 (19-25/1/2017)

5th iGEM Meeting
Location: Lab
Date: 20/2/2017
Time: 16:15-17:50
Attendance: All core members
In today’s meeting, we reported our research progress and started assembling our project.
We have also done an unofficial interview to the potential new iGEMers.

### Week 14 (26/2/2017-4/3/2017)

6th iGEM Meeting
Location: Lab
Date: 27/2/2017
Time: 16:15-18:00
Attendance: Most of the members
In today’s meeting, 7th graders showed us their STEM (Science, Technology, Engineering and Mathematics) project.
We were the judges in their competition. After the competition, core members of the team stayed behind to
discuss the further details of our iGEM trip.

### Week 15 (5-11/3/2017)

7th iGEM Meeting
Location: Lab
Date: 7/3/2017
Time: 16:15-17:45
Attendance:All of the members
We had a regular meeting today.

### Week 16 (12-18/3/2017)

Uh oh... we were lazy this week... wanna see what's happening next week?

### Week 17 (19-25/3/2017)

8th iGEM Meeting
Location: Lab
Date: 20/3/2017
Time: 16:00-17:30
Attendance: All of the members
Decided our research details, Human Practice, Collaboration details and division of labour

9th iGEM Meeting
Location: PMQ @ 35 Aberdeen Street, Central, Hong Kong
Date: 22/3/2017
Time: 19:00-21:30
Attendance: All of the members
We attended a talk organized by DIY Bio, a community lab in Hong Kong featuring Genome Editing.
The talk was given by Dr Alan S.L. Wong, an assistant professor of the School of the Biomedical Sciences
of the University of Hong Kong. We established a collaborative relationship with the community lab
and had received many pieces of precious advice from the seniors from the field in particular.

### Week 18 (26/3/2017-1/4/2017)

10th iGEM Meeting
Location: Lab
Date: 27/3/2017
Time: 16:00-17:30
Attendance: Most of the members
We had a meeting with four guest teachers from G. T. (Ellen Yeung) College. Sharing our experience of
participating in the 2016 iGEM, we hope to help them with their establishment of their new iGEM team.

11th iGEM Meeting
Location: Lab
Date: 30/3/2017
Time: 16:00-17:30
Attendance: Most of the members
We listed most pathways that produces uric acid, did some digging on those pathways, trying to come up
with a method to regulate the production of uric acid and we comb our data by writing the information we
have on hand on the windows in lab!

### Week 19 (2-8/4/2017)

12th iGEM Meeting
Location: HKUST
Date: 3/4/2017-4/4/2017
Time: 09:30-17:30
Attendance: All members
We attended another workshop called Regulatory issues associated with Genome Editing and Gene Drive -
Regulation of Synthetic Biology in Asia organized by professor K. L. Chow and his team from the Hong Kong
University of Science and Technology. Bioethics and more information about the latest genome editing technology,
CRISPR is given. We established collaborative relationships with HKUST and CUHK 2017 iGEM team respectively.
It was a fruitful workshop.

### Week 20 (9-15/4/2017)

Uh oh... we were lazy this week... wanna see what's happening in Week 24?

### Week 21 (16-22/3/2017)

Uh oh... we were lazy this week... wanna see what's happening in Week 24?

### Week 22 (23-29/4/2017)

Uh oh... we were lazy this week... wanna see what's happening in Week 24?

### Week 23 (30/4/2017-6/5/2017)

Uh oh... we were lazy this week... wanna see what's happening Week 24?

### Week 24 (7-13/5/2017)

13th iGEM Meeting
Location: Lab
Date: 8/5/2017
Time: 16:00-17:30
Attendance: Most of the members
We continued learning how to design parts and order it.

### Week 25 (14-20/5/2017)

Uh oh... we were lazy this week... wanna see what's happening in Week 27?

### Week 26 (21-27/5/2017)

Uh oh... we were lazy this week... wanna see what's happening in Week 27?

### Week 27 (28/5/2017-3/6/2017)

14th iGEM Meeting
Location: Lab
Date: 30/5/2017
Time: 15:20-17:30
Attendance: Most of the members
We have finalized the design of 3 parts.

### Week 28 (4-10/6/2017)

Uh oh... we were lazy this week... wanna see what's happening in Week 31?

### Week 29 (11-17/6/2017)

Uh oh... we were lazy this week... wanna see what's happening in Week 31?

### Week 30 (18-24/6/2017)

Uh oh... we were lazy this week... wanna see what's happening in Week 31?

### Week 31 (25/6/2017-1/7/2017)

15th iGEM Meeting
Location: Lab
Date 29/6/2017
Time: 13:15-15:00
Attendance: Most of our members
We had a short meeting after our exams to finalize our project progress and deadlines.
We are also briefed the guideline of the projects from iGEM

### Week 32 (2-8/7/2017)

16th iGEM Meeting
Location: Lab
Date: 4/7/2017
Time: 10:30-12:30
Attendance: Most of the members
Attendees were informed the blueprint of our wet lab schedule, they grasped an understanding on
the whole process of our phase 1 project. Protocols of the corresponding procedures were also combed.

17th iGEM Meeting
Location: Lab
Date: 5/7/2017
Time: 09:00-17:30
Attendance: Most of the members
Our phase 1 wet lab schedule was finalized and pre-lab calculations were done. New iGEMers had a bit
hands on wet lab training as well as presentation training. They tried to present our project once to the old iGEMers.
Throughout the process, they were taught some presentation skills and some conceptual mistakes were corrected.

### Week 33 (9-15/7/2017)

18th iGEM Meeting
Location: Lab
Date: 11/7/2017
Time: 15:00-17:00
Attendance: Most of the members
We prepared agar plate and SOB medium. Cells were seeded as well

19th iGEM Meeting
Location: Lab
Date: 12/7/2017
Time: 09:00-17:00
Attendance: Most of the members
We prepared competent cells, did restriction enzyme digestion, gel purification, ligation and transformation

20th iGEM Meeting
Location: Lab
Date: 13/7/2017
Time: 15:30-17:30
Attendance: Most of the members
We cloned our cells and did liquid medium culture

21st iGEM Meeting
Location: Lab
Date: 14/7/2017
Time: 10:00-12:00
Attendance: Most of the members
We did miniprep and stored cells for further experiments.

### Week 34 (16-22/7/2017)

22nd iGEM Meeting
Location: Lab
Date: 18/7/2017
Time: 14:00-16:30
Attendance: Some of the members
We prepared SOB medium and trained new iGEMers’ presentation skills.
Liquid culture and cell passing were also performed

23rd iGEM Meeting
Location: Lab
Date: 19/7/2017
Time: 10:00-12:30
Attendance: Some of the members
We did miniprep and gel electrophoresis of cloone checking. Training on presentation skills was done.

24th iGEM Meeting
Location: Lab
Date: 22/7/2017
Time: 10:00-12:30
Attendance: Most of the members
We organized the second meeting with iGEM 2017 Team CUHK to further discuss collaboration details.
We reached a consensus regarding the date and time borrowing their laboratory for our experiment as our
school laboratory is less well-equipped comparing to theirs. In return, we helped them build their ideal model
using 3D printing design software. We all had a fruitful meeting.

### Week 35 (23-29/7/2017)

25th iGEM Meeting
Location: Lab
Date: 25/7/2017
Time: 10:00-17:00
Attendance: Some of the members
We did restriction enzyme cut and gel electrophoresis today.

### Week 36 (30/7/2017-5/8/2017)

26th iGEM Meeting
Location: School Lab
Date: 1/8/2017
Time: 09:00-17:30
Attendance: Some of the members
We are honored to have the chance to perform our experiment in 2017 team CUHK laboratory. We once again did
restriction enzyme cut, gel purification, ligation, transformation and spread plate.

27th iGEM Meeting
Location: School Lab
Date: 2/8/2017
Time: 10:00-16:30
Attendance: Some of the members
We picked clone.

28th iGEM Meeting
Location: School Lab
Date: 3/8/2017
Time: 10:00-18:00
Attendance: Some of the members
We did miniprep, twice restriction enzyme cuts and gel electrophoresis.

29th iGEM Meeting\_Pan-Pearl iGEM Teams Meeting 2017
Location: Science Center LT1 in The Chinese University of Hong Kong
Date: 5/8/2017
Time: 10:30-17:30
Attendance: Some of the members
In the meeting we consulted representative from 2017 team Hong\_Kong\_HKU’s opinion as well as professional
information from prof. King-Lau Chow and King Ming CHAN.

### Week 37 (6-12/8/2017)

30th iGEM Meeting
Location: Lab
Date: 7/8/2017
Time: 14:00-17:00
Attendance: Some of the members
We did gel electrophoresis

### Week 38 (13-19/8/2017)

31st iGEM Meeting
Location: Lab
Date: 17/8/2017
Time: 09:00-17:30
Attendance: Some of the members
We did restriction enzyme cut, gel purification and ligation

32nd iGEM Meeting
Location: Lab
Date: 18/8/2017
Time: 10:00-14:00
Attendance: Some of the members
We did transformation and we have spreaded plate.

### Week 39 (20-26/8/2017)

33rd iGEM Meeting
Location: Lab
Date: 24/8/2017
Time: 09:00-17:30
Attendance: Some of the members
We did restriction enzyme cut, gel purification and ligation

34th iGEM Meeting
Location: Lab
Date: 25/8/2017
Time: 11:00-15:00
Attendance: Some of the members
We did transformation and we have spread plate.

35th iGEM Meeting
Location: Lab
Date: 26/8/2017
Time: 14:00-17:00
Attendance: Some of the members
We cloned our cells

36th iGEM Meeting
Location: Lab
Date: 28/8/2017
Time:14:00-15:30
Attendance: Some of our students
We picked clone.

### Week 40 (27/8/2017-2/9/2017)

37th iGEM Meeting
Location: Lab
Date: 29/8/2017
Time: 14:00-16:30
Attendance: Some of our students
We did miniprep.

38th iGEM Meeting
Location: Lab
Date: 30/8/2017
Time: 15:00-17:30
Attendance: Some of the members
We did assay test

### Week 41 (3-9/9/2017)

39th iGEM Meeting
Location: InnoSpace @ UCCKE
Date: 4/9/2017
Time: 16:30-18:00
Attendance: All core members
We had the very first meeting in this school year, Wiki page work allocation was conducted

### Week 42 (10-16/9/2017)

40th iGEM Meeting
Location: InnoSpace @ UCCKE
Date: 13/9/2017
Time: 13:00-15:00
Attendance: All core members
We discussed in details regarding the DSS STEM Fair we are going to take part in.
We have designed some games and booths for students as well as visitors to take part in.
iGEM will also be introduced to guests in order to raise their awareness regarding synthetic biology.

### Week 43 (17-23/9/2017)

41st iGEM Meeting
Location: Lab
Date: 17/9/2017
Time: 14:00-18:00
Attendance: Some of the members
We did transformation and have spread plates today.

42nd iGEM Meeting
Location: Lab
Date: 18/9/2017
Time: 17:30-20:00
Attendance: Some of the members
We cloned our cells.

### Week 44 (14-30/9/2017)

43rd iGEM Meeting\_DSS STEM Fair
Location: St. Paul Convent School
Date: 23-24/9/2017
Time: 09:00-18:00
Attendance: All members
DSS STEM Fair was organized in the favor of promoting STEM Education, which consists of
Science, Technology, Engineering and Mathematics. This fair is open to public and through
introducing to them our project along with the iGEM Community, it is hoped the awareness
among the public towards synthetic biology can be raised.

44th iGEM Meeting
Location: Lab
Date: 27/9/2017
Time: 14:30-18:00
Attendance: Some of the members
We did restriction enzyme cut, gel purification and ligation

45th iGEM Meeting
Location: Lab
Date: 28/9/2017
Time: 17:30-21:00
Attendance: Some of the members
We did transformation and have spread plate.

#### UCCKE iGEM 2017

#### Contact

* ucckeigem2017@gmail.com
* +852 2343 6677

#### Quicklinks

* [About](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Team)
* [Project](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Description)
* [Parts](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Parts)

#### Social Media

* [Facebook](https://www.facebook.com/ucckeigem2017/)
* [YouTube](https://www.youtube.com/channel/UCNXwNbYHbTisrHdccfkW5hQ)

UCCKE team. Designed with  by Gabriel Lee.

[![](https://static.igem.org/mediawiki/2017/b/bf/HQ_menu_logo.jpg)](https://2017.igem.org)


[![](https://static.igem.org/mediawiki/2017/8/8b/HQ_page_logo.jpg)](https://2017.igem.org)

# Team:Hong Kong UCCKE/Experiments

Toggle navigation     [UCCKE 2017](https://2017.igem.org/Team%3AHong_Kong_UCCKE)

* About
  + [Team](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Team)
  + [Collaborations](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Collaborations)
  + [Contact](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Contact)
* Project
  + [Description](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Description)
  + [Design](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Design)
  + [Experiments](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Experiments)
  + [Results](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Results)
  + [Contribution](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Contribution)
  + [Notebook](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Notebook)
  + [Protocols](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Protocols)
  + [Attributions](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Attributions)
* Parts
  + [Parts](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Parts)
  + [Basic Parts](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Basic_Part)
  + [Composite Parts](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Composite_Part)
* [Safety](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Safety)
* Human Practices
  + [Silver HP](https://2017.igem.org/Team%3AHong_Kong_UCCKE/HP/Silver)
  + [Public Engagement](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Engagement)
* [Judging Form](https://igem.org/2017_Judging_Form?team=Hong_Kong_UCCKE)

## Experiments

We have done 4 assays, which are Assay A, Assay B, Assay C and Assay D on the cultured cells to test whether part BBa\_K2197300, BBa\_K2197400, BBa\_K2197500 and BBa\_K2197502 are valid. Protocols are documented on the protocol page. The design and results will be discussed in detail below.

### Assay A

Design

Assay A is an experiment designed to prove the validity of BBa\_K2197300. By incubating engineered cells with different concentrations of uric acid and measuring the green fluorescence level in a plate reader, it is expected a positively proportional trend be shown. As theoretically, the higher the concentration is, the more the GFP will be expressed. It is because the strong repressor KRAB-HucR will stop the expression of GFP when there is no uric acid, or in very low concentration.

Steps and Result

We first pipette the cells into different concentrations of uric acid and incubate them in 96 well plate for 30 minutes. After that, we put it into the plate reader to measure the green fluorescence level.

The result is shown as below (Click me for raw data).

[![Adding uric acid to the cell](https://static.igem.org/mediawiki/2017/f/fc/T--Hong_Kong_UCCKE--300graph.jpg)](https://static.igem.org/mediawiki/2017/f/fc/T--Hong_Kong_UCCKE--300graph.jpg "Adding uric acid to the cell")

There is no significant trend shown. When higher concentrations of uric acid are added, the levels of green fluorescence measured are similar to that in lower concentrations. Therefore, we carry out another experiment, Assay B, after receiving the advice from 2017 team Hong Kong-CUHK.

[![Assay A](https://static.igem.org/mediawiki/2017/3/3b/T--Hong_Kong_UCCKE--uricacid.jpg)](https://static.igem.org/mediawiki/2017/3/3b/T--Hong_Kong_UCCKE--uricacid.jpg "Assay A")Assay A
[![Assay A](https://static.igem.org/mediawiki/2017/e/e0/T--Hong_Kong_UCCKE--96wellplate.jpg)](https://static.igem.org/mediawiki/2017/e/e0/T--Hong_Kong_UCCKE--96wellplate.jpg "Assay A")Assay A

### Assay B

Design

Assay B is another experiment designed to prove the validity of BBa\_K2197300. By growing engineered cells with different concentrations of uric acid and measuring the green fluorescence level in a plate reader, we expect a positively proportional trend will be shown. As theoretically, the higher the concentration is, the more the GFP will be expressed. It is because the strong repressor KRAB-HucR will stop the expression of GFP when there is no uric acid, or in very low concentration.

Steps and Result

We first grew cells in LB broth overnight. Then we cultured cells from the same tube with different concentrations of uric acid-LB mixed. We then incubate the cells for 1 hour and then pipette them into the 96-well plate. After that, we put it into the plate reader to measure the green fluorescence level.

The result is shown as below (Click to see raw data).

[![Growing the cell with uric acid](https://static.igem.org/mediawiki/2017/2/21/T--Hong_Kong_UCCKE--300wifcellgraph.jpg)](https://static.igem.org/mediawiki/2017/2/21/T--Hong_Kong_UCCKE--300wifcellgraph.jpg "Growing the cell with uric acid")

There is also no significant trend shown. When higher concentrations of uric acid are added, the amount of GFP measured is similar to that in lower concentration. Therefore, we tried to figure out the problem. After double checking the sequence, we found out that the DNA sequence we ordered from IDT is different from what we designed, in which the sequence 'tactagag' is missing. GFP E0040, Terminator B0010, K2197302 and K2197303 are affected. Referring to the mechanism of the promoter, if HucO is missing, the repressor protein mUTS can not bind to it. Thus, the amount of GFP expression cannot be restricted and no trend can be shown. Also, the GFP is seriously affected, therefore, GFP cannot be expressed.

[![Assay B](https://static.igem.org/mediawiki/2017/9/91/T--Hong_Kong_UCCKE--cellsgrowinuricacid-LBmixed.jpg)](https://static.igem.org/mediawiki/2017/9/91/T--Hong_Kong_UCCKE--cellsgrowinuricacid-LBmixed.jpg "Assay B")Assay B

### Assay C

Design

Assay C is an experiment designed to prove the validity of BBa\_K2197400. By incubating engineered cells (BBa\_K2197400) and Competent cells with different concentrations of uric acid, then adding engineered cells (BBa\_K2197300) after 30 minutes, we measure the Green fluorescence level in a plate reader. We expect that the columns with engineered cells (BBa\_K2197400) will have less green fluorescence expressed when comparing to the column containing competent cells. As theoretically, the higher the concentration is, the more the smUOX will be expressed and this will catalyse uric acid into allantoin. So by adding engineered cells (BBa\_K2197300) which expresses more GFP when there is more uric acid, the green fluorescence level of the columns with engineered cells(BBa\_K2197400) will be lower than the columns with competent cells although they were in the same concentration at the beginning

Steps and Result

We first pipette the cells (BBa\_2197400, competent cells) into different concentrations of uric acid and incubate them in 96 well plate for 30 minutes. After that, we add in engineered cells (BBa\_K2197300) into the wells and incubate them for another 30 minutes. Then, we put it into the plate reader for the measurement of green fluorescence level.

However, due to the problem of BBa\_K2197300, in which there is no difference of the green fluorescence levels between the one with lower uric acid concentration and higher uric acid concentration, we cannot determine whether it can really lower the level of uric acid.

### Assay D

Design

Assay D is an experiment designed to prove the validity of BBa\_K2197500 and BBa\_K2197502. By incubating engineered cells (BBa\_K2197500), cells (BBa\_K2197502) and Competent cells with different concentrations of uric acid, then adding engineered cells (BBa\_K2197300) after 30 minutes, we measure the Green fluorescence level in a plate reader. We expect that the columns with engineered cells (BBa\_K2197500) and engineered cells (BBa\_K2197502) will have less green fluorescence expressed when comparing to the column containing competent cells. As theoretically, engineered cells (BBa\_K2197500) will absorb uric acid into the cell and for engineered cells (BBa\_K2197502), the higher the concentration is, the more uric acid will be absorbed into the cells. So by adding engineered cells (BBa\_K2197300) which express more GFP when there is more uric acid, the green fluorescence level of the columns with engineered cells (BBa\_K2197500 and BBa\_K2197502) will be lower than the columns with competent cells although they were in the same concentration at the beginning.

Steps and Result

We first pipette the cells (BBa\_2197500, BBa\_K2197502, competent cells) into different concentrations of uric acid and incubate them in 96 well plate for 30 minutes. After that, we add engineered cells (BBa\_K2197300) into the wells and incubate for another 30 minutes. Then, we put it into the plate reader for the measurement of green fluorescence level.

However, due to the problem of BBa\_K2197300, in which there is no difference of the green fluorescence levels between the one with lower uric acid concentration and higher uric acid concentration, we cannot determine whether it can really lower the level of uric acid.

#### UCCKE iGEM 2017

#### Contact

* ucckeigem2017@gmail.com
* +852 2343 6677

#### Quicklinks

* [About](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Team)
* [Project](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Description)
* [Parts](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Parts)

#### Social Media

* [Facebook](https://www.facebook.com/ucckeigem2017/)
* [YouTube](https://www.youtube.com/channel/UCNXwNbYHbTisrHdccfkW5hQ)

UCCKE team. Designed with  by Gabriel Lee.

[![](https://static.igem.org/mediawiki/2017/b/bf/HQ_menu_logo.jpg)](https://2017.igem.org)

[![](https://static.igem.org/mediawiki/2017/8/8b/HQ_page_logo.jpg)](https://2017.igem.org)

# Team:Hong Kong UCCKE/Results

Toggle navigation     [UCCKE 2017](https://2017.igem.org/Team%3AHong_Kong_UCCKE)

* About
  + [Team](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Team)
  + [Collaborations](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Collaborations)
  + [Contact](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Contact)
* Project
  + [Description](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Description)
  + [Design](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Design)
  + [Experiments](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Experiments)
  + [Results](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Results)
  + [Contribution](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Contribution)
  + [Notebook](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Notebook)
  + [Protocols](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Protocols)
  + [Attributions](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Attributions)
* Parts
  + [Parts](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Parts)
  + [Basic Parts](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Basic_Part)
  + [Composite Parts](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Composite_Part)
* [Safety](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Safety)
* Human Practices
  + [Silver HP](https://2017.igem.org/Team%3AHong_Kong_UCCKE/HP/Silver)
  + [Public Engagement](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Engagement)
* [Judging Form](https://igem.org/2017_Judging_Form?team=Hong_Kong_UCCKE)

## Results

After we have successfully cloned our plasmids into the E.coli, we did miniprep to obtain purified plasmid DNA. Then, we did restriction map and sent the samples to BGI for sequencing. The results are shown below.

### BBa\_K2197300

Restriction Map

After mini prep, we used the same restriction enzymes EcoR1 and Pst1 to do restriction digestion and gel electrophoresis. However, the size of K2197300 and the backbone pSB1C3 are almost the same, thus we cannot prove by the Gel photo.

[![Image of gel electrophoresis of 300a+b](https://static.igem.org/mediawiki/2017/8/82/T--Hong_Kong_UCCKE--theoracpic300.jpg)](https://static.igem.org/mediawiki/2017/8/82/T--Hong_Kong_UCCKE--theoracpic300.jpg "Image of gel electrophoresis of 300a+b")
Image of gel electrophoresis of 300a+b

Sequencing Result

With the help of team 2017 Hong\_Kong-CUHK, we sent our plasmid to BGI for sequencing. We aligned the sequence we ordered from IDT (As the template sequence 99593011 below) with the sequencing result received from BGI (as the aligned sequence below). And here is our analysis.

The black area indicates the two sequences are the same which those red strips represent there're some mutations.

[![300 sequencing resultl](https://static.igem.org/mediawiki/2017/c/c6/T--Hong_Kong_UCCKE--300sequence.jpg)](https://static.igem.org/mediawiki/2017/c/c6/T--Hong_Kong_UCCKE--300sequence.jpg "300 sequencing result")

Sequencing result of BBa\_K2197300

[![](https://static.igem.org/mediawiki/2017/1/1b/T--Hong_Kong_UCCKE--_project_300_screenshot_1.png)](https://static.igem.org/mediawiki/2017/1/1b/T--Hong_Kong_UCCKE--_project_300_screenshot_1.png "Allignment (1)")

Sequence Alignment(1)

[![](https://static.igem.org/mediawiki/2017/1/16/T--Hong_Kong_UCCKE--_project_300_screenshot_2.png)](https://static.igem.org/mediawiki/2017/1/16/T--Hong_Kong_UCCKE--_project_300_screenshot_2.png "Project 300 (2)")

Sequence Alignment(2)

Conclusion

From the results above, we can say we have successfully cloned the DNA that IDT sent us into the cell. However, as what we have mentioned in the experiment page, we found that the cells cannot express GFP. Then, we aligned the sequence registered in iGEM with the one that IDT send us, we found that the sequence containing 'tactagag' is missing.

[![](https://static.igem.org/mediawiki/2017/c/ca/T--Hong_Kong_UCCKE--sequenceofigem2.jpg)](https://static.igem.org/mediawiki/2017/c/ca/T--Hong_Kong_UCCKE--sequenceofigem2.jpg "Alignment (1)")

Sequence Alignment(1)

[![](https://static.igem.org/mediawiki/2017/7/75/T--Hong_Kong_UCCKE--sequenceofigem1.jpg)](https://static.igem.org/mediawiki/2017/7/75/T--Hong_Kong_UCCKE--sequenceofigem1.jpg "Alignment (2)")

Sequence Alignment(2)

### BBa\_K2197400

Restriction Map

After mini prep, we used the same restriction enzymes EcoR1 and Pst1 to do restriction digestion and gel electrophoresis. As seen, the position of K2197400 (2214 bp) and the backbone pSB1C3 (2070 bp) are similar to our predicted result.

[![](https://static.igem.org/mediawiki/2017/5/54/T--Hong_Kong_UCCKE--21asdfghj2.jpg)](https://static.igem.org/mediawiki/2017/5/54/T--Hong_Kong_UCCKE--21asdfghj2.jpg "Image of gel electrophoresis of 500")

Image of gel electrophoresis of 400a+b and 500

[![](https://static.igem.org/mediawiki/2017/2/2b/T--Hong_Kong_UCCKE--theoracpic400.jpg)](https://static.igem.org/mediawiki/2017/2/2b/T--Hong_Kong_UCCKE--theoracpic400.jpg "Image of gel electrophoresis of 500")

Image of theoretical gel electrophoresis of 400 from genome compiler

Sequencing Result

With the help of team 2017 Hong\_Kong-CUHK, we sent our plasmid to BGI for sequencing. We aligned the sequence we ordered from IDT (As the template sequence 163014570 below) with the sequencing result received from BGI (as the aligned sequence below). And here is our analysis.

The black area indicates that the two sequences are the same which those red strips represent there're some mutations.

![](https://static.igem.org/mediawiki/2017/c/c1/T--Hong_Kong_UCCKE--gelsequence400.png)

Sequencing Result of BBa\_K2197400

[![](https://static.igem.org/mediawiki/2017/2/2d/T--Hong_Kong_UCCKE--_project_400_screenshot_1.png)](https://static.igem.org/mediawiki/2017/2/2d/T--Hong_Kong_UCCKE--_project_400_screenshot_1.png "Project 400 (1)")

Sequence Alignment (1)

[![](https://static.igem.org/mediawiki/2017/e/e1/T--Hong_Kong_UCCKE--_project_400_screenshot_2.png)](https://static.igem.org/mediawiki/2017/e/e1/T--Hong_Kong_UCCKE--_project_400_screenshot_2.png "Project 400 (2)")

Sequence Alignment (2)

Conclusion

From the results above, we can say we have successfully cloned the plasmid DNA into the cell.

### BBa\_K2197500

#### Restriction Map

Also using the same restriction enzymes EcoR1 and Pst1 to do restriction digestion and gel electrophoresis. The position of K2197500 (1668 bp) and the backbone pSB1C3 (2070 bp) is similar to our predicted result.

[![](https://static.igem.org/mediawiki/2017/d/dc/T--Hong_Kong_UCCKE--gelpic400ab500.jpeg)](https://static.igem.org/mediawiki/2017/d/dc/T--Hong_Kong_UCCKE--gelpic400ab500.jpeg "Image of gel electrophoresis of 500")

Image of gel electrophoresis of 400a+b and 500

[![](https://static.igem.org/mediawiki/2017/d/de/T--Hong_Kong_UCCKE--sgelofi500.jpg)](https://static.igem.org/mediawiki/2017/d/de/T--Hong_Kong_UCCKE--sgelofi500.jpg "Image of gel electrophoresis of 500")

Image of theoretical gel electrophoresis of 500 from genome compiler

Sequencing Result

We sent our plasmid to BGI for sequencing. We aligned the sequence we ordered from IDT (As the template sequence 163014569 below) with the sequencing result received from BGI(as the aligned sequence below). And here is our analysis.

The black area indicates that the two sequences are the same which those red strips represent that there're some mutations.

[![Project 500](https://static.igem.org/mediawiki/2017/f/fd/T--Hong_Kong_UCCKE--500abcdefghijklmnoph.jpg)](https://static.igem.org/mediawiki/2017/f/fd/T--Hong_Kong_UCCKE--500abcdefghijklmnoph.jpg "Project 500")
Sequencing Result of BBa-K2197500

[![](https://static.igem.org/mediawiki/2017/6/63/T--Hong_Kong_UCCKE--_project_500_screenshot_1.png)](https://static.igem.org/mediawiki/2017/6/63/T--Hong_Kong_UCCKE--_project_500_screenshot_1.png "Project 500 (1)")

Project 500 (1)

[![](https://static.igem.org/mediawiki/2017/b/be/T--Hong_Kong_UCCKE--_project_500_screenshot_2.png)](https://static.igem.org/mediawiki/2017/b/be/T--Hong_Kong_UCCKE--_project_500_screenshot_2.png "Project 500 (2)")

Project 500 (2)

Conclusion

From the results above, we can say we have successfully cloned the plasmid DNA into the cell.

### BBa\_K2197502

Sequencing Result

We sent our plasmid to BGI for sequencing. We aligned the sequence that we ordered from IDT (As the template sequence 163014567 below) with the sequencing result received from BGI (as the aligned sequence below). And here is our analysis.

The black area indicates that the two sequences are the same which those red stripsrepresent that there're some mutations.

[![502 sequencing resultl](https://static.igem.org/mediawiki/2017/3/32/T--Hong_Kong_UCCKE--sequenceofi500.jpg)](https://static.igem.org/mediawiki/2017/3/32/T--Hong_Kong_UCCKE--sequenceofi500.jpg "502 sequencing result")

Sequencing result of BBa\_K2197502

[![](https://static.igem.org/mediawiki/2017/4/48/T--Hong_Kong_UCCKE--_project_502_screenshot_1.png)](https://static.igem.org/mediawiki/2017/4/48/T--Hong_Kong_UCCKE--_project_502_screenshot_1.png "Project 502 (1)")

Project 502 (1)

[![](https://static.igem.org/mediawiki/2017/4/40/T--Hong_Kong_UCCKE--_project_502_screenshot_2.png)](https://static.igem.org/mediawiki/2017/4/40/T--Hong_Kong_UCCKE--_project_502_screenshot_2.png "Project 502 (2)")

Project 502 (2)

Conclusion

From the results above, we can say we have successfully cloned the plasmid DNA into the cell.

#### UCCKE iGEM 2017

#### Contact

* ucckeigem2017@gmail.com
* +852 2343 6677

#### Quicklinks

* [About](https://2017.igem.org/Team%3AHong_Kong_UCCKE/Team)
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