BioBrick Workshop

iGEM Denmark 2013.

We planned and hosted a three-days BioBrick workshop to introduce ourselves and the other iGEM teams from Denmark (SDU, KU) to the lab skills necessary to complete our projects. The workshop consisted of a mix of lectures, lab work and social events.

During the lab work.

Lectures were on the BioBrick standards, BioBricks registry, project and time management, and the methods used over the lab experiments of the workshop, such as: USER cloning, PCR reactions and purification of plasmids, etc.

The purpose of this workshop was to served as an assembly tutorial that covered skills independent of organism and project. And of course to have fun!!

Creative use of pipettes!

Classic case of photobombing!

Project

We will combine 5 parts using USER cloning in E. coli, and will validate that the transformation was successful.

Agenda

Preparation (Done at DTU, by DTU team earlier in the week) Prepare plates (4 per team) Prepare colonies to use for validating that the transformation has worked

Friday March 8th Location: 208/62

13:00 -- Welcome, Introductions (Chris Workman)

13:30 -- Lecture: Introduction to working with Biobricks (Thomas Trolle) Discussion of the Biobrick standard Overview of the lab process

14:30 -- Lecture: What is the biobrick registry, and how to find parts (Damian Plichta) Work through an example of finding parts to reach a particular goal.

15:00 -- Lecture: Introduction to the weekend project and USER cloning (Mathilde Lund) What is USER cloning? USER cloning with Biobricks

16:30 -- Lecture: PHUSER (Hans Jasper Genee)

17:00 -- Tour of the lab Introduction to the equipment and techniques that will be over the weekend

18:00 -- Social dinner in CBS kitchen

Saturday March 9th Location:

9:00 -- Introduction to labwork (Mathilde Lund) Safety considerations when working in the lab. How to run PCR

10:30 -- Begin work on the project 1. Prepare PCR reaction mix (2 PCR reaction pr team) SOP1 2. Prepare USER cloning reaction mix SOP 3 + handout note (1 positive and 1 negative reaction per team) Step 1 and 2 can both be stored several hours at 4°C

12:00 -- Lunch (Pizza)

13:00 -- Lecture: Tips and Tricks in the lab (TBD) Practical advice from previous iGEMmers on what techniques worked well; what pitfalls they encountered, and how to avoid them.

13:30 -- Lecture: Advice on project management and time management (Julie Rank)

14:00 -- Continue work on project in the lab (Transform cells and plate, incubate overnight)

1. Prepare 1 analytical gel for each team + 2 purification gels SOP2 2. 2. While waiting for the gel, prepare samples for analytical gel + purification electrophorese SOP2 3. Prepare transformation of USER cloning SOP3 4. While waiting for the transformation: Load analytical gel + purification gel electrophorese SOP2 4. Heatshock of transformation and plating of transformation SOP3 5. Analyse analytical gel by UV SOP2 6. Cut DNA from purification gel; freeze at -18°C for next day or purify right away SOP2

20:00 -- Social Dinner at India Palace (buffet) Adress: 13 H.C. Andersens Boulevard 1553 Copenhagen V

Sunday March 10th Location:

9:00 -- Continue work on project in the lab (Using pre-prepared cultures, digest and run on a gel) 1. Purify plasmids SOP4 2. Prepare restriction enzyme mix for purified plasmids SOP4 3. Analyse transformation plate and count number of colonies of transformation plates from Saturday. Pick 3 colonies from positive plate for further cultivation and purification SOP4

12:00 -- Lunch (Pizza?)

13:00 -- Continue Lab work 4. Purify DNA from cutout of gel from Saturday SOP2

13:30 -- Lecture: How to get gold in iGEM (Chris Workman) Discuss some of the frequently overlooked aspects of iGEM such as human practices and characterization of Biobricks that are necessary to achieve a gold medal.

14:30 -- Finish lab work

5. Prepare restriction enzyme reaction samples for gel electrophoresis SOP2 6. Load on gel SOP2

16:00 -- Wrap up, Thanks (Chris Workman)