Troubleshooting

• Introduction
• General Tips and Tricks
1. Transformation
• iGEM Protocols
• Answers
• Ask Your Questions

Welcome to the Troubleshooting page!

My name is Traci, and I'm here to help you with your cloning problems.

You can contact me by email at traci AT igem DOT org, on Reddit Traci_at_iGEM, or on Twitter @Traci_Haddock

Introduction

Cloning is difficult. No one who has picked up a pipette in the lab would debate this fact. However, cloning shouldn't be impossible! This page exists to help teams who are struggling with cloning and other molecular techniques. Treat this page as another resource in your troubleshooting endeavors rather than the be-all, end-all resource for iGEM cloning problems. This is an experimental project for the 2015 iGEM season and I hope this will be a helpful resource for every iGEM team!

As the Science and Technology Fellow at iGEM Headquarters, I want to try to help you out with cloning problems to the best of my abilities. To this end, I will be offering advice and possible solutions to your problems. As I start collecting questions and generating answers, I plan to create a Troubleshooting Guide that will serve as a resource for both current and future teams to use throughout their iGEM experience.

My background is primarily in bacterial cloning with a focus on working in E. coli. I completed my doctorate in cell and molecular biology from the University of Rhode Island in 2011 and was a postdoctoral researcher in synthetic biology with Dr. Douglas Densmore at Boston University from 2011-2013. From there, I continued to work in the lab as the Executive Director of the Boston University Center of Synthetic Biology and I joined the iGEM Headquarters team in April 2015. In summary, I have over 14 years of experience with molecular techniques and cloning in E. coli, and over 4 years of experience with synthetic biology in particular.

General Tips and Tricks

This section is meant to provide some general advice for cloning problems. I recommend you read through this material if you're having problems and/or while you're waiting for a response to your specific question. Each link below is geared towards a specific area where cloning problems occur. If you have other tips you'd like me to include in these pages, please send them using the question form below!

• E. coli Transformations



iGEM Protocols

Below are the links to the various protocols that iGEM Headquarters has provided over the years. These protocols can also be found through the Parts Registry page under the "Help" and "Protocols" links in the black toolbar at the top of the page.

These are another useful resource that teams should read through when they're experiencing cloning problems.



• 3A assembly kit
• Transformation efficiency kit
• PCR standardization
• Linearized plasmid backbone
• Transformation
• Miniprep
• Restriction digest
• Ligation
• Making competent cells
• Antibiotic stocks



Answers to Your Questions

As teams start to submit their questions, I'll be posting answers here.

I'll also make sure to Tweet whenever I update this page, so please follow me on Twitter: @Traci_Haddock




Ask Your Questions Here!

First and foremost, there are no stupid questions, especially when it comes to cloning problems. Ask me literally anything about cloning! I'm also happy to answer questions about general molecular biology techniques (including plasmid minipreps, DNA gel electrophoresis, and so on).

Your name, team, and email will be kept confidential, but I will repost your question when I answer it. I'm asking that everyone provide a first name and an email so I can follow-up with you if your question is unclear. Emails will come from traci at igem dot org if I need to follow-up with you.

Thanks very much for participating!