iGEM 2025 — Team hkssc

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{% block title %}Notebook{% endblock %}
{% block lead %}This serves as a chronological record of your team's progress throughout the season. It documents your daily activities, experiments, discussions, and decisions.{% endblock %}
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# The General Timeline for Our Main Project

## Dec.

### Research Starts

Confirm topic: Eczema

## March

### Confirm genes of interest

FLG, HSP70, fish collagen

## April

### Consultation from CUHK and HKU

Narrow gene of interest to FLG

## May

### The First Engineering Cycle Starts

## June

### CUHK skin model workshop

## July

### Lab work within school

## August

### The Second Engineering Cycle Starts

Lab work within school

## Sep.

### Lab work ends

Lab work within school and HKU

# Project Planning

Initial project proposal(March)

We proposed that combined treatment of these proteins with artificial FLG in coconut oil could restore the expression of FLG while promoting its stability and overall relieving AD symptoms. We emailed our proposal to HKU and CUHK for further advice.

CUHK meeting(8/4)

With their guidance, we narrowed down the three proposed proteins—FLG, HSP70, and fish collagen—to FLG alone, as the other two presented challenges: HSP70 is widely studied in other laboratories, and fish collagen proved too complex for our current capabilities. Additionally, they suggested that a skin model, transwell assay, and scratch assay could serve as pivotal methodologies for our experiments.

HKU meeting(28/4)

This has allowed us to refine our project ideas, including creating disease models in cells by using inflammatory-inducing genes (eg LPS, IL4, IL13) to trigger inflammatory response, instead of our original plan of using FLG knocked out cells, which we found out is not only difficult to execute, but also how it doesn’t really equate to AD cells. Furthermore, they also enriched our knowledge on FLG, NMF and experiment procedures on protein expression and extraction (IPTG induction and SDS Page etc.).

# Project Lab Work

|  |  |  |  |  |
| --- | --- | --- | --- | --- |
| Day   Count | Date | Work | Link to protocol | Notes |
| Day 1 | 13/5 | Media preparation | [13/5 Day1 Media Preparation](https://www.google.com/url?q=https://docs.google.com/document/d/12n26ke3XUA22nN-qq5qwbXAWqVwKnsLxJ8G65bluSDY/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503097944&usg=AOvVaw2QfiaJE6ou1yeBucwNGWgr) |  |
| Day 2 | 14/5 | Hydration of plasmid | [14/5 Day2 Hydrating plasmid](https://www.google.com/url?q=https://docs.google.com/document/d/1RHVFkpwdST2DdcIm8NHZ7Qd9VqLbe9E6a3yPIKqlKnE/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503098695&usg=AOvVaw2xJd-Hua_c5vAxgAg_v16f) |  |
| Day 3 | 15/5 | Transformation and plating | [15/5 Day3 Transformation](https://www.google.com/url?q=https://docs.google.com/document/d/1RjoR6UmdA27n32x5knLnw-I-fcatzvS0TTT0oHkhbfQ/edit&sa=D&source=editors&ust=1759343503099318&usg=AOvVaw1EQlcOquzwjcF25zETVg5b) |  |
| Day 4 | 16/5 | Review transformation results | [16/5 Day4 transformation results fail](https://www.google.com/url?q=https://docs.google.com/document/d/1RjslTe8F_iMqGMyT9jKbeJiVwWj1GRRm6YjCvI5M2eg/edit&sa=D&source=editors&ust=1759343503100000&usg=AOvVaw0Gq0sm-xt0FWOu7H_ts-ao) | Fail, no colonies observed |
| Day 5 | 19/5 | Transformation | [19/5 Day5 Transformation](https://www.google.com/url?q=https://docs.google.com/document/d/1EbPHZICbeAYlLBoRH0HzSUL6bblywjlhnAMG1SePyp8/edit&sa=D&source=editors&ust=1759343503100576&usg=AOvVaw1fFAkygC7Sj-5xXvPMnug9) | Modification: 45 seconds for heat shock |
| Day 6 | 20/5 | Review transformation results | [20/5 Day6 Transformation results success](https://www.google.com/url?q=https://docs.google.com/document/d/1RmweyHTPk19DWyL4hvR435hg6TgwebLNP0nBOFOH-jk/edit&sa=D&source=editors&ust=1759343503101180&usg=AOvVaw2n5M31700zUJm1AXJHwSWE) | Success, although one negative control is contaminated |
| Day 6 | 20/5 | Growing single bacteria colony | [20/5 Day6 Single colony inoculation](https://www.google.com/url?q=https://docs.google.com/document/d/1Oy1eIa3J3ZYY9449RC1F7AF1CFMzDZw-URcutABldGM/edit&sa=D&source=editors&ust=1759343503101788&usg=AOvVaw3x0Ai3D71rHwcN7qB5YNKz) |  |
| Day 7 | 21/5 | Plasmid isolation | [21/5 Day7 Plasmid isolation](https://www.google.com/url?q=https://docs.google.com/document/d/18VqXh9OQT_emUZ3eIc5rag_fq0yhu2CElyMRFHVf4-0/edit&sa=D&source=editors&ust=1759343503102330&usg=AOvVaw09C0m7NAaFTUr-9KMP4x1T) | No.1,3,4 were dropped and spilled |
| Day 7 | 21/5 | Nanodrop | [21/5 pET-iDT-FLG nanodrop](https://www.google.com/url?q=https://docs.google.com/spreadsheets/d/10bkZg2PD205wXurJR-VT-GH_urK3ncdiaPPrestynqo/edit&sa=D&source=editors&ust=1759343503102922&usg=AOvVaw0goidNAysKEIrrIeUxH4iG) | All the sample concentration are very low |
| Day 8 | 26/5 | Inoculation | [26/5 Day8 Single colony inoculation](https://www.google.com/url?q=https://docs.google.com/document/d/1_KGsM_Y1u02tgpG_H6bxQdl74w33MlP1mdiztUHLx1U/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503103628&usg=AOvVaw3oIpJMydIaSnAYeme7BAf-) |  |
| Day 9 | 27/5 | Plasmid isolation and nanodrop | [27/5 Day9 Plasmid isolation](https://www.google.com/url?q=https://docs.google.com/document/d/1lmO8wj6JohfE_Pcb-cvT70h-xYTJOgw12XNPcrRYyjo/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503104203&usg=AOvVaw34bK3z4Z4boBqX7n7RODMq) | Highest concentration: 113ng/uL |
| Day 10 | 28/5 | Restriction digestion and gel electrophoresis | [28/5 Day10 Restriction digestion](https://www.google.com/url?q=https://docs.google.com/document/d/1Dek2o5YpNpHwXg6YxYFycfHfdhARwyfhIvaxSMyrunw/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503104841&usg=AOvVaw1D-LLYb5ggeyizxxLLA9ng) |  |
| Day 11 | 25/6 | Media | [25/6 Day11 Media preparation](https://www.google.com/url?q=https://docs.google.com/document/d/1TjXgSM_Pd8-lqiXw6zLijkRafb-VsbWxXAws-lEoSuQ/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503105453&usg=AOvVaw1Bh5HsSE4tAHr2VwCVv4np) |  |
| Day 12 | 26/6 | Workshop at CUHK | [26/6 CUHK Workshop Day0 (Helen)](https://www.google.com/url?q=https://docs.google.com/document/d/17gJ7RV85ltwtLF5EiqhhdsluYrSWLObsgjWzwGom1uA/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503106100&usg=AOvVaw2p1FXxkCfQYOhDTXNO27jS) | Seeded cells and did scratch assay |
| Day 13 | 27/6 | Workshop at CUHK | [27/6 CUHK Workshop Day2 (Helen)](https://www.google.com/url?q=https://docs.google.com/document/d/18Vacg7Ytnik6OKGvByZee5mWRgB20CDipc9nekZlVGI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503106721&usg=AOvVaw3Tx2s1g7gvkwYl4fE_c6C8)  [27/6 iGEM CUHK workshop (Evelyn)](https://www.google.com/url?q=https://docs.google.com/document/d/1ojXxjXh4rgO7R6lsSrCIvrNMxkuAVDIvpgySiy6qzng/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503106937&usg=AOvVaw0EarztJZr_2LzR1N2dY6dB) |  |
| Day 14 | 3/7 | Transformation | [3/7 Day14 Transformation](https://www.google.com/url?q=https://docs.google.com/document/d/1q9YReF-6XsFbbHoW0Malu4hCWmvvQHSAHJ2ofqVFrDs/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503107629&usg=AOvVaw1JsnMwQf0NAvJjhUTF62dx) |  |
| Day 15 | 10/7 | ColonyPCR preparation | [10/7 Day15 ColonyPCR preparation](https://www.google.com/url?q=https://docs.google.com/document/d/1HbX9q25nLlX9KaTCvgDZI0wwbyY20tJxre6rkPyhQpQ/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503108285&usg=AOvVaw2Dpjfi0VcCwJJmfCwtiORp) |  |
| Day 16 | 14/7 | ColonyPCR | [14/7 Day16 ColonyPCR](https://www.google.com/url?q=https://docs.google.com/document/d/1SM7l1aG_yrUM6AfcV3KwU8DMmZOAqjIGDb7o5E1RBXM/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503108881&usg=AOvVaw2sJ9M5qu__4Twq7hmz7jF6) |  |
| Day 17 | 15/7 | Gel electrophoresis and colonyPCR | [15/7 Day17 Gel electrophoresis and single colony inoculation](https://www.google.com/url?q=https://docs.google.com/document/d/1JYU_SYSgvSb_47KpKAXQT-UV2u7qMvasc93XD8WC4s8/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503109477&usg=AOvVaw3JAD3u2FTwizrUcoUDJg3t) | No bands observed, failed |
| Day 18 | 16/7 | Gel electrophoresis |  |  |
| Day 19 | 17/7 | IPTG induction | [17/7 Day19 IPTG Protein expression.docx](https://www.google.com/url?q=https://docs.google.com/file/d/1gVzXPQ8PITOQzhw-xgqTaYjQERYNLr59/edit?usp%3Ddocslist_api%26filetype%3Dmsword&sa=D&source=editors&ust=1759343503110722&usg=AOvVaw117QpI3LiyY8g35K2ASk73) |  |
| Day 20 | 18/7 | Cell harvesting |  |  |
| Day 21 | 22/7 | Protein extraction | [22/7 Day21 Sample prepartion for SDS PAGE-1.docx](https://www.google.com/url?q=https://docs.google.com/file/d/1lztoC51gRcl96-BqMZjC8JjyTFYHt0SK/edit?usp%3Ddocslist_api%26filetype%3Dmsword&sa=D&source=editors&ust=1759343503111886&usg=AOvVaw1Tv0o008Ua92gnTSqAbywI) |  |
| Day 22 | 23/7 | colonyPCR and cell harvesting | [23/7 Day22 Protein expression \_250718\_125956.docx](https://www.google.com/url?q=https://docs.google.com/file/d/125_zfFUBhZJ3P2RLw_M2KGodt5qB8NAu/edit?usp%3Ddocslist_api%26filetype%3Dmsword&sa=D&source=editors&ust=1759343503112496&usg=AOvVaw0iyX7BujLcHuD4Q3YwQs41)  [23/7 Day22 ColonyPCR](https://www.google.com/url?q=https://docs.google.com/document/d/1rCDm-flWdbhEWnQLKe5qtRaZr_4xicL3hhoPMt5SvaQ/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503112683&usg=AOvVaw2FVx1P79FQ5XZJH7rRxySB) |  |
| Day 23 | 24/7 | SDS-PAGE |  |  |
| Day 24 | 25/7 | Sample preparation for SDS PAGE for 0.5mM | [25/7 Day24 Sample prepartion for SDS PAGE-1.docx](https://www.google.com/url?q=https://docs.google.com/file/d/1y3ML3kVUGiq6_9df2wRK5U2Wa3W3dY5W/edit?usp%3Ddocslist_api%26filetype%3Dmsword&sa=D&source=editors&ust=1759343503113824&usg=AOvVaw1Y43UEmWuLKBs3Zu4vn_t3) |  |
| Day 25 | 28/7 | SDS-PAGE for 0.5mM |  |  |
| Day 26 | 30/7 | Sample preparation for 1mM SDS-PAGE | [30/7 Day26 Sample prepartion for SDS PAGE-1.docx](https://www.google.com/url?q=https://docs.google.com/file/d/1K3D2Czj-YMqA_q9vjkcD1fYEQ1JOrHiE/edit?usp%3Ddocslist_api%26filetype%3Dmsword&sa=D&source=editors&ust=1759343503114929&usg=AOvVaw3lgpmsEGKj15G8z1Hadtqm) |  |
| Day 27 | 31/7 | Destaining of SDS PAGE 1mM and Bacteria growth curve |  | Dont know how to use the spectrophotometer |
| Day 28 | 1/8 | Bacteria growth curve at37°C | [1/8 Bacterial growth curve](https://www.google.com/url?q=https://docs.google.com/spreadsheets/d/17IBDN8L1sLMx_-JR8qFErzRQ7DdM-2eBiG0GxPMOlqk/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503116051&usg=AOvVaw35nmb01dhZ0MDImm9YDnUk) |  |
| Day 29 | 4/8 | Bacteria growth curve at 37°C | [1/8 Bacterial growth curve](https://www.google.com/url?q=https://docs.google.com/spreadsheets/d/17IBDN8L1sLMx_-JR8qFErzRQ7DdM-2eBiG0GxPMOlqk/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503116630&usg=AOvVaw0GFBsWZ1f2pbOKNKzuFhwI) |  |
| Day 30 | 6/8 | Bacteria growth curve at 37°C |  |  |
| Day 31 | 7/8 | Bacteria growth curve at 37°C and  Cell lysate | [7/8 cell lysate protocol + notes](https://www.google.com/url?q=https://docs.google.com/document/d/1FwM8ygfmnYAHy6g8I6nTyLHCie6_XxNt7x-CzJMdzk4/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503117888&usg=AOvVaw1DDJQ64lMjDSvJ1cUSesWl) |  |
| Day 32 | 8/8 | Bacteria growth curve at 37°C |  |  |
| Day 33 | 11/8 | preparing primary culture, FLG purification at RT, cell lysate |  |  |
| Day 34 | 12/8 | IPTG expression at 20°C, bacterial growth curve, FLG purification at 37°C | [12/8 FLG purification nano drop](https://www.google.com/url?q=https://docs.google.com/spreadsheets/d/10OjtVN99L1BhECSQi1mmzmE53i8pgZwVKuLsy63ou9I/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503119505&usg=AOvVaw0-kafo5J6JtoPxtCnMMN5n) | [12/8 bacterial growth curve 20°C](https://www.google.com/url?q=https://docs.google.com/spreadsheets/d/10Z0_iGxzWZQ0mPimv--n7vUgxHuc2HiVkaqaZseqTQQ/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503119692&usg=AOvVaw2_tZerpx8pVeETFoxqjo_6) |
| Day 35 | 13/8 | IPTG expression at 20°C, preparing FLG at 37°C for SDS-PAGE | [13/8 Day35 Protein samples for SDS-PAGE.docx](https://www.google.com/url?q=https://docs.google.com/file/d/1YRbnwDBJKwbTqhCv1PLRML3K2_i4B-6G/edit?usp%3Ddocslist_api%26filetype%3Dmsword&sa=D&source=editors&ust=1759343503120282&usg=AOvVaw2T_09bR2tMd-dYYz6ZPLlz) | [12/8 FLG purification nano drop](https://www.google.com/url?q=https://docs.google.com/spreadsheets/d/10OjtVN99L1BhECSQi1mmzmE53i8pgZwVKuLsy63ou9I/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503120521&usg=AOvVaw1DuZKgxc0y6Rl42JvqVLic) |
| Day 36 | 15/8 |  |  |  |
| Day 37 | 18/8 | SDS-PAGE for total protein at 20°C |  |  |
| Day 38 | 19/8 | Destaining of SDS-PAGE gel |  |  |
| Day 39 | 20/8 | Dry lab work |  |  |
| Day 40 | 21/8 | Dry lab work |  |  |
| Day 41 | 25/8 | HKU lab  Seeding cells | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503124379&usg=AOvVaw3PO2ubfdPcLisWhuAmYA8b) |  |
| Day 42 | 27/8 | HKU lab preparing stock, working concentration and wells | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503125064&usg=AOvVaw2Ng7hKAw08US-FNgitGRiv) |  |
| Day 43 | 28/8 | HKU lab harvesting cells at T24 | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503125737&usg=AOvVaw3TkYeuXDKsz-ykBhf0wJVO) |  |
| Day 44 | 29/8 | HKU lab harvesting cells at T48 | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503126367&usg=AOvVaw37nqW3_4r4UshtCnurHf2t) |  |
| Day 45 | 2/9 | HKU lab  Extracting RNA T1 | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503127090&usg=AOvVaw11SKxFZA9szcFWAoKtFVa7) |  |
| Day 46 | 3/9 | CYCLE 2 START  Media preparation 200ml agar, 10 plates |  |  |
| Day 47 | 4/9 | Transformation and plating | [4/9 Day47 Transformation](https://www.google.com/url?q=https://docs.google.com/document/d/15B7nB_RtLxv7cBLkVU_vcs8BBqIIymBvuHzNbd3ANgg/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503128323&usg=AOvVaw1th9eLwgVdPLMWvr5XReok) |  |
| Day 48 | 5/9 | HKU lab  Extracting RNA T24 and T48 | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503128997&usg=AOvVaw3bhK1vgRyFWC5Ypc-qNw_a) |  |
| Day 49 | 9/9 | HKU lab reverse transcription of all samples | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503129629&usg=AOvVaw0H2F_rlpZ_xXMzB6F2JLyG) |  |
| Day 50 | 10/9 | Colony PCR for  idT-pET-FLG-RFP | [8/9 Day49 ColonyPCR](https://www.google.com/url?q=https://docs.google.com/document/d/1hBQh5i50FtnJRdivBrJPRNEBqLQSBIQCJMtWKU_UD58/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503130210&usg=AOvVaw1wSPDt_DTpBQjhKxGSflCV) |  |
| Day 51 | 11/9 | Gel electrophoresis |  |  |
| Day 52 | 16/9 | Single colony inoculation | [16/9 Day52 Single colony inoculation](https://www.google.com/url?q=https://docs.google.com/document/d/1hD_8oj6H7MLvPprG-NO1BRA8eofE2L_AhVMTwdOIYvk/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503131384&usg=AOvVaw3G97cj5qp-H02S6lVF6v6F) |  |
| Day 53 | 17/9 | Plasmid isolation + nanodrop | [17/9 pET-FLG-RFP plasmid isolation nano drop results](https://www.google.com/url?q=https://docs.google.com/spreadsheets/d/11NGUZ_KDT-CeMYg5niOAhCY01XeCncjj385KgtvwXu0/edit?usp%3Dsharing&sa=D&source=editors&ust=1759343503132113&usg=AOvVaw3F13EwI_kYfADJf0YXs7Df) |  |
| Day 54 | 18/9 | HKU qPCR | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503132721&usg=AOvVaw1CGBNI0lvMGqrxBvcqbNC3) |  |
| Day 55 | 19/9 | RD for confirmation of clone | [19/9 Day53 Restriction digestion](https://www.google.com/url?q=https://docs.google.com/document/d/1p7cHb968Bw-i6Xd2VzHXWP45QQ_y1aGCa_VloNQQQe4/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503133350&usg=AOvVaw11JCWkLVgn_NhBjeus3ybt) | Failed again ah;( |

{% endblock %}

{% extends "layout.html" %}
{% block title %}Engineering Success{% endblock %}
{% block lead %}Demonstrate engineering success in a technical aspect of your project by going through at least one iteration of the engineering design cycle.{% endblock %}
{% block prop %}

{% endblock %}
{% block page\_content %}{% endblock %}

{% extends "layout.html" %}
{% block title %}Experiments{% endblock %}
{% block lead %}Describe the research, experiments, and protocols you used in your project. It is designed to provide sufficient information for other teams to replicate our work.{% endblock %}
{% block prop %}

{% endblock %}
{% block page\_content %}

![](https://static.igem.wiki/teams/5613/oth/a.webp)

# Cycle 1

### Wet lab workflow in SSC

1. Design the Plasmid FLG ✅
2. Clone the plasmid in E.coli DH5alpha✅
3. Single bacterial colony inoculation✅
4. Plasmid isolation followed by NanoDrop✅- Confirmation of clone by restriction digestion✅
   - Running gel electrophoresis✅
   - Sub cloning the plasmid into E.coli BL21 ✅
   - Confirmation of clone by colony PCR ✅
   - Single colony inoculation✅
   - IPTG induction and protein expression✅
   - Running the SDS protein gel✅
   - Protein extraction✅
   - Plotting bacterial growth curve against the protein expression.✅

### Workshop in CUHK

1. SCRATCH assay using heat shock protein and collagen✅

### Wet lab in HKU

1. qPCR for Carbonic anhydrase II (CA2), IL-24, ribosomal protein L38 (RPL38) and FLG ✅

# Cycle 2

1. Design the Plasmid FLG✅
2. Clone the plasmid in E.coli DH5alpha✅
3. Colony PCR for the confirmation of clone😞
4. Gel electrophoresis
5. Plasmid isolation
6. Clone the plasmid into BL21
7. Colony PCR for the confirmation of clone
8. Gel electrophoresis

|  |  |  |  |  |
| --- | --- | --- | --- | --- |
| Day   Count | Date | Work | Link to protocol | Notes |
| Day 1 | 13/5 | Media preparation | [13/5 Day1 Media Preparation](https://www.google.com/url?q=https://docs.google.com/document/d/12n26ke3XUA22nN-qq5qwbXAWqVwKnsLxJ8G65bluSDY/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503097944&usg=AOvVaw2QfiaJE6ou1yeBucwNGWgr) |  |
| Day 2 | 14/5 | Hydration of plasmid | [14/5 Day2 Hydrating plasmid](https://www.google.com/url?q=https://docs.google.com/document/d/1RHVFkpwdST2DdcIm8NHZ7Qd9VqLbe9E6a3yPIKqlKnE/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503098695&usg=AOvVaw2xJd-Hua_c5vAxgAg_v16f) |  |
| Day 3 | 15/5 | Transformation and plating | [15/5 Day3 Transformation](https://www.google.com/url?q=https://docs.google.com/document/d/1RjoR6UmdA27n32x5knLnw-I-fcatzvS0TTT0oHkhbfQ/edit&sa=D&source=editors&ust=1759343503099318&usg=AOvVaw1EQlcOquzwjcF25zETVg5b) |  |
| Day 4 | 16/5 | Review transformation results | [16/5 Day4 transformation results fail](https://www.google.com/url?q=https://docs.google.com/document/d/1RjslTe8F_iMqGMyT9jKbeJiVwWj1GRRm6YjCvI5M2eg/edit&sa=D&source=editors&ust=1759343503100000&usg=AOvVaw0Gq0sm-xt0FWOu7H_ts-ao) | Fail, no colonies observed |
| Day 5 | 19/5 | Transformation | [19/5 Day5 Transformation](https://www.google.com/url?q=https://docs.google.com/document/d/1EbPHZICbeAYlLBoRH0HzSUL6bblywjlhnAMG1SePyp8/edit&sa=D&source=editors&ust=1759343503100576&usg=AOvVaw1fFAkygC7Sj-5xXvPMnug9) | Modification: 45 seconds for heat shock |
| Day 6 | 20/5 | Review transformation results | [20/5 Day6 Transformation results success](https://www.google.com/url?q=https://docs.google.com/document/d/1RmweyHTPk19DWyL4hvR435hg6TgwebLNP0nBOFOH-jk/edit&sa=D&source=editors&ust=1759343503101180&usg=AOvVaw2n5M31700zUJm1AXJHwSWE) | Success, although one negative control is contaminated |
| Day 6 | 20/5 | Growing single bacteria colony | [20/5 Day6 Single colony inoculation](https://www.google.com/url?q=https://docs.google.com/document/d/1Oy1eIa3J3ZYY9449RC1F7AF1CFMzDZw-URcutABldGM/edit&sa=D&source=editors&ust=1759343503101788&usg=AOvVaw3x0Ai3D71rHwcN7qB5YNKz) |  |
| Day 7 | 21/5 | Plasmid isolation | [21/5 Day7 Plasmid isolation](https://www.google.com/url?q=https://docs.google.com/document/d/18VqXh9OQT_emUZ3eIc5rag_fq0yhu2CElyMRFHVf4-0/edit&sa=D&source=editors&ust=1759343503102330&usg=AOvVaw09C0m7NAaFTUr-9KMP4x1T) | No.1,3,4 were dropped and spilled |
| Day 7 | 21/5 | Nanodrop | [21/5 pET-iDT-FLG nanodrop](https://www.google.com/url?q=https://docs.google.com/spreadsheets/d/10bkZg2PD205wXurJR-VT-GH_urK3ncdiaPPrestynqo/edit&sa=D&source=editors&ust=1759343503102922&usg=AOvVaw0goidNAysKEIrrIeUxH4iG) | All the sample concentration are very low |
| Day 8 | 26/5 | Inoculation | [26/5 Day8 Single colony inoculation](https://www.google.com/url?q=https://docs.google.com/document/d/1_KGsM_Y1u02tgpG_H6bxQdl74w33MlP1mdiztUHLx1U/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503103628&usg=AOvVaw3oIpJMydIaSnAYeme7BAf-) |  |
| Day 9 | 27/5 | Plasmid isolation and nanodrop | [27/5 Day9 Plasmid isolation](https://www.google.com/url?q=https://docs.google.com/document/d/1lmO8wj6JohfE_Pcb-cvT70h-xYTJOgw12XNPcrRYyjo/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503104203&usg=AOvVaw34bK3z4Z4boBqX7n7RODMq) | Highest concentration: 113ng/uL |
| Day 10 | 28/5 | Restriction digestion and gel electrophoresis | [28/5 Day10 Restriction digestion](https://www.google.com/url?q=https://docs.google.com/document/d/1Dek2o5YpNpHwXg6YxYFycfHfdhARwyfhIvaxSMyrunw/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503104841&usg=AOvVaw1D-LLYb5ggeyizxxLLA9ng) |  |
| Day 11 | 25/6 | Media | [25/6 Day11 Media preparation](https://www.google.com/url?q=https://docs.google.com/document/d/1TjXgSM_Pd8-lqiXw6zLijkRafb-VsbWxXAws-lEoSuQ/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503105453&usg=AOvVaw1Bh5HsSE4tAHr2VwCVv4np) |  |
| Day 12 | 26/6 | Workshop at CUHK | [26/6 CUHK Workshop Day0 (Helen)](https://www.google.com/url?q=https://docs.google.com/document/d/17gJ7RV85ltwtLF5EiqhhdsluYrSWLObsgjWzwGom1uA/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503106100&usg=AOvVaw2p1FXxkCfQYOhDTXNO27jS) | Seeded cells and did scratch assay |
| Day 13 | 27/6 | Workshop at CUHK | [27/6 CUHK Workshop Day2 (Helen)](https://www.google.com/url?q=https://docs.google.com/document/d/18Vacg7Ytnik6OKGvByZee5mWRgB20CDipc9nekZlVGI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503106721&usg=AOvVaw3Tx2s1g7gvkwYl4fE_c6C8)  [27/6 iGEM CUHK workshop (Evelyn)](https://www.google.com/url?q=https://docs.google.com/document/d/1ojXxjXh4rgO7R6lsSrCIvrNMxkuAVDIvpgySiy6qzng/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503106937&usg=AOvVaw0EarztJZr_2LzR1N2dY6dB) |  |
| Day 14 | 3/7 | Transformation | [3/7 Day14 Transformation](https://www.google.com/url?q=https://docs.google.com/document/d/1q9YReF-6XsFbbHoW0Malu4hCWmvvQHSAHJ2ofqVFrDs/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503107629&usg=AOvVaw1JsnMwQf0NAvJjhUTF62dx) |  |
| Day 15 | 10/7 | ColonyPCR preparation | [10/7 Day15 ColonyPCR preparation](https://www.google.com/url?q=https://docs.google.com/document/d/1HbX9q25nLlX9KaTCvgDZI0wwbyY20tJxre6rkPyhQpQ/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503108285&usg=AOvVaw2Dpjfi0VcCwJJmfCwtiORp) |  |
| Day 16 | 14/7 | ColonyPCR | [14/7 Day16 ColonyPCR](https://www.google.com/url?q=https://docs.google.com/document/d/1SM7l1aG_yrUM6AfcV3KwU8DMmZOAqjIGDb7o5E1RBXM/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503108881&usg=AOvVaw2sJ9M5qu__4Twq7hmz7jF6) |  |
| Day 17 | 15/7 | Gel electrophoresis and colonyPCR | [15/7 Day17 Gel electrophoresis and single colony inoculation](https://www.google.com/url?q=https://docs.google.com/document/d/1JYU_SYSgvSb_47KpKAXQT-UV2u7qMvasc93XD8WC4s8/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503109477&usg=AOvVaw3JAD3u2FTwizrUcoUDJg3t) | No bands observed, failed |
| Day 18 | 16/7 | Gel electrophoresis |  |  |
| Day 19 | 17/7 | IPTG induction | [17/7 Day19 IPTG Protein expression.docx](https://www.google.com/url?q=https://docs.google.com/file/d/1gVzXPQ8PITOQzhw-xgqTaYjQERYNLr59/edit?usp%3Ddocslist_api%26filetype%3Dmsword&sa=D&source=editors&ust=1759343503110722&usg=AOvVaw117QpI3LiyY8g35K2ASk73) |  |
| Day 20 | 18/7 | Cell harvesting |  |  |
| Day 21 | 22/7 | Protein extraction | [22/7 Day21 Sample prepartion for SDS PAGE-1.docx](https://www.google.com/url?q=https://docs.google.com/file/d/1lztoC51gRcl96-BqMZjC8JjyTFYHt0SK/edit?usp%3Ddocslist_api%26filetype%3Dmsword&sa=D&source=editors&ust=1759343503111886&usg=AOvVaw1Tv0o008Ua92gnTSqAbywI) |  |
| Day 22 | 23/7 | colonyPCR and cell harvesting | [23/7 Day22 Protein expression \_250718\_125956.docx](https://www.google.com/url?q=https://docs.google.com/file/d/125_zfFUBhZJ3P2RLw_M2KGodt5qB8NAu/edit?usp%3Ddocslist_api%26filetype%3Dmsword&sa=D&source=editors&ust=1759343503112496&usg=AOvVaw0iyX7BujLcHuD4Q3YwQs41)  [23/7 Day22 ColonyPCR](https://www.google.com/url?q=https://docs.google.com/document/d/1rCDm-flWdbhEWnQLKe5qtRaZr_4xicL3hhoPMt5SvaQ/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503112683&usg=AOvVaw2FVx1P79FQ5XZJH7rRxySB) |  |
| Day 23 | 24/7 | SDS-PAGE |  |  |
| Day 24 | 25/7 | Sample preparation for SDS PAGE for 0.5mM | [25/7 Day24 Sample prepartion for SDS PAGE-1.docx](https://www.google.com/url?q=https://docs.google.com/file/d/1y3ML3kVUGiq6_9df2wRK5U2Wa3W3dY5W/edit?usp%3Ddocslist_api%26filetype%3Dmsword&sa=D&source=editors&ust=1759343503113824&usg=AOvVaw1Y43UEmWuLKBs3Zu4vn_t3) |  |
| Day 25 | 28/7 | SDS-PAGE for 0.5mM |  |  |
| Day 26 | 30/7 | Sample preparation for 1mM SDS-PAGE | [30/7 Day26 Sample prepartion for SDS PAGE-1.docx](https://www.google.com/url?q=https://docs.google.com/file/d/1K3D2Czj-YMqA_q9vjkcD1fYEQ1JOrHiE/edit?usp%3Ddocslist_api%26filetype%3Dmsword&sa=D&source=editors&ust=1759343503114929&usg=AOvVaw3lgpmsEGKj15G8z1Hadtqm) |  |
| Day 27 | 31/7 | Destaining of SDS PAGE 1mM and Bacteria growth curve |  | Dont know how to use the spectrophotometer |
| Day 28 | 1/8 | Bacteria growth curve at37°C | [1/8 Bacterial growth curve](https://www.google.com/url?q=https://docs.google.com/spreadsheets/d/17IBDN8L1sLMx_-JR8qFErzRQ7DdM-2eBiG0GxPMOlqk/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503116051&usg=AOvVaw35nmb01dhZ0MDImm9YDnUk) |  |
| Day 29 | 4/8 | Bacteria growth curve at 37°C | [1/8 Bacterial growth curve](https://www.google.com/url?q=https://docs.google.com/spreadsheets/d/17IBDN8L1sLMx_-JR8qFErzRQ7DdM-2eBiG0GxPMOlqk/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503116630&usg=AOvVaw0GFBsWZ1f2pbOKNKzuFhwI) |  |
| Day 30 | 6/8 | Bacteria growth curve at 37°C |  |  |
| Day 31 | 7/8 | Bacteria growth curve at 37°C and  Cell lysate | [7/8 cell lysate protocol + notes](https://www.google.com/url?q=https://docs.google.com/document/d/1FwM8ygfmnYAHy6g8I6nTyLHCie6_XxNt7x-CzJMdzk4/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503117888&usg=AOvVaw1DDJQ64lMjDSvJ1cUSesWl) |  |
| Day 32 | 8/8 | Bacteria growth curve at 37°C |  |  |
| Day 33 | 11/8 | preparing primary culture, FLG purification at RT, cell lysate |  |  |
| Day 34 | 12/8 | IPTG expression at 20°C, bacterial growth curve, FLG purification at 37°C | [12/8 FLG purification nano drop](https://www.google.com/url?q=https://docs.google.com/spreadsheets/d/10OjtVN99L1BhECSQi1mmzmE53i8pgZwVKuLsy63ou9I/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503119505&usg=AOvVaw0-kafo5J6JtoPxtCnMMN5n) | [12/8 bacterial growth curve 20°C](https://www.google.com/url?q=https://docs.google.com/spreadsheets/d/10Z0_iGxzWZQ0mPimv--n7vUgxHuc2HiVkaqaZseqTQQ/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503119692&usg=AOvVaw2_tZerpx8pVeETFoxqjo_6) |
| Day 35 | 13/8 | IPTG expression at 20°C, preparing FLG at 37°C for SDS-PAGE | [13/8 Day35 Protein samples for SDS-PAGE.docx](https://www.google.com/url?q=https://docs.google.com/file/d/1YRbnwDBJKwbTqhCv1PLRML3K2_i4B-6G/edit?usp%3Ddocslist_api%26filetype%3Dmsword&sa=D&source=editors&ust=1759343503120282&usg=AOvVaw2T_09bR2tMd-dYYz6ZPLlz) | [12/8 FLG purification nano drop](https://www.google.com/url?q=https://docs.google.com/spreadsheets/d/10OjtVN99L1BhECSQi1mmzmE53i8pgZwVKuLsy63ou9I/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503120521&usg=AOvVaw1DuZKgxc0y6Rl42JvqVLic) |
| Day 36 | 15/8 |  |  |  |
| Day 37 | 18/8 | SDS-PAGE for total protein at 20°C |  |  |
| Day 38 | 19/8 | Destaining of SDS-PAGE gel |  |  |
| Day 39 | 20/8 | Dry lab work |  |  |
| Day 40 | 21/8 | Dry lab work |  |  |
| Day 41 | 25/8 | HKU lab  Seeding cells | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503124379&usg=AOvVaw3PO2ubfdPcLisWhuAmYA8b) |  |
| Day 42 | 27/8 | HKU lab preparing stock, working concentration and wells | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503125064&usg=AOvVaw2Ng7hKAw08US-FNgitGRiv) |  |
| Day 43 | 28/8 | HKU lab harvesting cells at T24 | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503125737&usg=AOvVaw3TkYeuXDKsz-ykBhf0wJVO) |  |
| Day 44 | 29/8 | HKU lab harvesting cells at T48 | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503126367&usg=AOvVaw37nqW3_4r4UshtCnurHf2t) |  |
| Day 45 | 2/9 | HKU lab  Extracting RNA T1 | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503127090&usg=AOvVaw11SKxFZA9szcFWAoKtFVa7) |  |
| Day 46 | 3/9 | CYCLE 2 START  Media preparation 200ml agar, 10 plates |  |  |
| Day 47 | 4/9 | Transformation and plating | [4/9 Day47 Transformation](https://www.google.com/url?q=https://docs.google.com/document/d/15B7nB_RtLxv7cBLkVU_vcs8BBqIIymBvuHzNbd3ANgg/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503128323&usg=AOvVaw1th9eLwgVdPLMWvr5XReok) |  |
| Day 48 | 5/9 | HKU lab  Extracting RNA T24 and T48 | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503128997&usg=AOvVaw3bhK1vgRyFWC5Ypc-qNw_a) |  |
| Day 49 | 9/9 | HKU lab reverse transcription of all samples | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503129629&usg=AOvVaw0H2F_rlpZ_xXMzB6F2JLyG) |  |
| Day 50 | 10/9 | Colony PCR for  idT-pET-FLG-RFP | [8/9 Day49 ColonyPCR](https://www.google.com/url?q=https://docs.google.com/document/d/1hBQh5i50FtnJRdivBrJPRNEBqLQSBIQCJMtWKU_UD58/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503130210&usg=AOvVaw1wSPDt_DTpBQjhKxGSflCV) |  |
| Day 51 | 11/9 | Gel electrophoresis |  |  |
| Day 52 | 16/9 | Single colony inoculation | [16/9 Day52 Single colony inoculation](https://www.google.com/url?q=https://docs.google.com/document/d/1hD_8oj6H7MLvPprG-NO1BRA8eofE2L_AhVMTwdOIYvk/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503131384&usg=AOvVaw3G97cj5qp-H02S6lVF6v6F) |  |
| Day 53 | 17/9 | Plasmid isolation + nanodrop | [17/9 pET-FLG-RFP plasmid isolation nano drop results](https://www.google.com/url?q=https://docs.google.com/spreadsheets/d/11NGUZ_KDT-CeMYg5niOAhCY01XeCncjj385KgtvwXu0/edit?usp%3Dsharing&sa=D&source=editors&ust=1759343503132113&usg=AOvVaw3F13EwI_kYfADJf0YXs7Df) |  |
| Day 54 | 18/9 | HKU qPCR | [HKU lab](https://www.google.com/url?q=https://docs.google.com/document/d/140KouMHbRVVrBB0kSi0tLC5HdPOdtp4hKi6Gd2DwYKI/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503132721&usg=AOvVaw1CGBNI0lvMGqrxBvcqbNC3) |  |
| Day 55 | 19/9 | RD for confirmation of clone | [19/9 Day53 Restriction digestion](https://www.google.com/url?q=https://docs.google.com/document/d/1p7cHb968Bw-i6Xd2VzHXWP45QQ_y1aGCa_VloNQQQe4/edit?usp%3Ddrivesdk&sa=D&source=editors&ust=1759343503133350&usg=AOvVaw11JCWkLVgn_NhBjeus3ybt) | Failed again ah;( |

{% endblock %}

{% extends "layout.html" %}
{% block title %}Parts{% endblock %}
{% block lead %}Present the results of your project, along with a detailed analysis and discussion of their significance. Also outline future plans and reflections on the impact of your project.{% endblock %}
{% block prop %}

{% endblock %}
{% block page\_content %}

## Results & Parts Characteristaion

---

|  |  |  |  |  |
| --- | --- | --- | --- | --- |
| Parts | Name | Type | Function | Results |
| [BBa\_250JDL7B](https://www.google.com/url?q=https://registry.igem.org/parts/bba-250jdl7b&sa=D&source=editors&ust=1758640370802121&usg=AOvVaw2gyeLl9Q77P17u0yT8y1HR) | FLG | Coding gene | The filaggrin (FLG) is a major epidermal protein that ensures the correct formations and function of the skin barrier. FLG can be broken down into natural moisturizing factors(NMF), including urocanic acid (UCA) and pyrrolidone carboxylic acid (PCA), which keeps the skin hydrated. [1] | Confirmed in DH5α, B(DE3)pLysS strains and fibroblast cells. |
| [BBa\_25EPAOF5](https://www.google.com/url?q=https://registry.igem.org/parts/bba-25epaof5&sa=D&source=editors&ust=1758640370804099&usg=AOvVaw2Rg7IIy_ZO4rNCRHPftuUN) | FLGxsc | Coding gene | The filaggrin (FLG) is a major epidermal protein that ensures the correct formations and function of the skin barrier. FLG can be broken down into natural moisturizing factors(NMF), including urocanic acid (UCA) and pyrrolidone carboxylic acid (PCA), which keeps the skin hydrated. [1] | Confirmed in DH5α cells. |
| [BBa\_J97003](https://www.google.com/url?q=https://parts.igem.org/Part:BBa_J97003&sa=D&source=editors&ust=1758640370805899&usg=AOvVaw3Tu50dEmQbkugUA0-W7j0l) | RFP | Coding gene | Fluorescent proteins (FPs) allow the specific and targeted labeling of proteins, organelles, cells, tissues, and whole organisms. This versatility makes FPs superior over synthetic organic dyes for imaging applications in vivo. [2] | N/A |
| [BBa\_J176131](https://www.google.com/url?q=https://parts.igem.org/Part:BBa_J176131&sa=D&source=editors&ust=1758640370807742&usg=AOvVaw11V3_IHOLP2UVL9uJOdwJc) | PLrigid | Protein domain | PLrigid is a 20 a.a. peptide that is based on a previously characterized alpha-helix motif (EAAAR) [3] | N/A |
| [BBa\_K2141000](https://www.google.com/url?q=https://parts.igem.org/Part:BBa_K2141000&sa=D&source=editors&ust=1758640370809221&usg=AOvVaw0tvKYYWThITqMs4J0sj4f_) | pET-28a(+) | Plasmid backbone | pET28a(+) is a nonviral expression plasmid used in bacteria strains for protein production purposes. It has kanamycin resistance gene as a selection marker. Plasmid sequence consists of T7 promoter and terminator sequences as well as lac operon sequence and lacI promoter for protein expression induction. It has both N- and C- terminal HisTag sequences and N- terminal thrombin cleavage site. | Confirmed in DH5α and B(DE3)pLysS strains |

---

**References**

[1]<https://pmc.ncbi.nlm.nih.gov/articles/PMC2721001/>

[2]<https://pmc.ncbi.nlm.nih.gov/articles/PMC4433748/>

[3]<https://pubmed.ncbi.nlm.nih.gov/2021618/>

{% endblock %}