Team:Newcastle/14 June 2010

This is the first day of the iGEM lab training. We were given several important tips by Dr. Wendy Smith to begin with, and then throughout the day, we were familiarized with basic lab techniques, e.g. the use of pipettes.

Wet Lab techniques practice

Aims

The aim of today's Lab training session was to practice aseptic techniques and wet lab techniques such as streak plating and broth culture preparation.

Materials Required

For today's session we needed:

• Agar plates
• Pipettes
• Wire loops
• E.coli (from a colony)
• LB broth
• Bunsen burner
• Conical flasks
• Orbital shaker

List of techniques

1. Made broth culture
2. Familiarised with using pipettes of different sizes
3. Mini-Prep introduction for Tuesday.
4. Used balance and made LB broth.

The biobrick BBa_J04450's prefix and suffix were identified. They are EcoRI and PstI respectively.

Tips

• Use aseptic technique: Treat everything as a pathogen!
• Work around the bunsen burner.
• Use a control.
• Clean the bench at the end of the day!
• Heat the wire loop from the middle to the tip.
• Keep plates upside down.
• Keep lids off for as short a time as possible.
• Loosen all the tops off the vessels before you start.
• Flame tops of bottles lightly.
• Colony plates should be labelled (name of culture, our initial, date) at the base in order to prevent condensation.
• For individual clony extraction, steak across a few times in different directions in order to dilute
• Wear gloves, especially when working with DNA, PCR etc. However, never wear gloves when working near a flame

Streak plating

Set up culture for minipreps

5 ml culture with E.coli from a single colony grown over night. Tomorrow we will harvest the DNA.

Outcome

We learnt aseptic technique and some basic wetlab techniques. As you can see in the picture above our streak plates worked.