Team:Newcastle/21 June 2010

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{{Team:Newcastle/mainbanner}}
{{Team:Newcastle/mainbanner}}
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'''Monday'''
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This is the '''FIRST DAY''' of the iGEM lab training for the second group. 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. how to use pipettes.
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This is the first day of the iGEM lab training. We were given several important tips by Wendy to begin with, and then throughout the day, we had to familiarise with basic lab techniques, e.g. the use of pipettes.
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=Wet Lab techniques practice=
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==Wet Lab techniques practice==
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===Aims===  
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==Aims==
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The aim of today's Lab training session was to practice aseptic technique and wet lab techniques such as streak plating and broth culture.  
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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.
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===Equipement list===
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==Materials Required==
'''For today's session we needed''':
'''For today's session we needed''':
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* ''E.coli'' (from a colony)  
* ''E.coli'' (from a colony)  
* LB broth  
* LB broth  
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* Bunsen burner  
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* Burnsen burner  
* Conical flasks
* Conical flasks
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* Orbital shaker  
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* Orbital shaker
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===List of techniques===
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==List of techniques==
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#Made broth culture
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#Used electronic balance and made LB broth.
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#Familiarised with using pipettes of different sizes
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#Prepared broth culture
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#Familiarized with using pipettes of different volumes
#Mini-Prep introduction for Tuesday.
#Mini-Prep introduction for Tuesday.
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#Used balance and made LB broth.
 
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The biobrick '''BBa_J04450''''s prefix and suffix were identified. They are EcoRI and PstI respectively.
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The biobrick '''BBa_J04450''''s prefix and suffix were identified from the parts registry. They are EcoRI and PstI respectively.
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===Tips===  
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==Tips==
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* Use Aseptic technique: Treat everything as a pathogen!
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* Use aseptic technique: Treat everything as a pathogen!
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* Work around the bunsen burner.
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* Work around burnsen burner.
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* Use a control.
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* Use a control for every experiment.
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* Clean the bench at the end of the day!
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* Clean the bench at the end of the day.
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* Heat the wire loop from the middle to the tip.  
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* Heat the flame loop from the middle to the tip till it becomes red hot.  
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* Keep plates upside down.  
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* Keep plates upside down so that  condensation does not damage the colonies.  
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* Keep lids off for as short a time as possible.
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* Keep the lids of the plates off for as short time as possible.
* Loosen all the tops off the vessels before you start.  
* Loosen all the tops off the vessels before you start.  
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* Flame tops of bottles lightly.
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* Flame tops of the bottles lightly so as to reduce the chances of contamination.
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* Colony plates should be labelled (name of culture, our initial, date) at the base in order to prevent condensation.
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* Colony plates should be labelled (name of culture, antibiotics used, our initials and date) at the base and inverted in order to prevent condensation.
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* For individual clony extraction, steak across a few times in different directions in order to dilute
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* For individual colony selection, steak across a few times in different directions in order to dilute the number of colonies per unit area.
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* Wear gloves, especially when working with DNA, PCR etc. However, never wear gloves when working near a flame
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* Wear gloves, especially when working with DNA, PCR etc. However, never wear gloves when working near a flame.
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===Streak plating===
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==Streak plating==
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[[Image:Newcastle_Deena_streakplate.JPG|alt="|Star plate streaking student, Deena, shows off how it is to be done!"|Caption="Star plate streaking student, Deena, shows off how it is to be done!"]]
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[[Image:Newcastle_Deena_streakplate.JPG|250px|Star plate-streaking student, Deena, shows how it is to be done!]]
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Star plate streaking student, Deena, shows off how it is to be done!
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'''Picture 1''': Picture showing Star plate-streaking student Deena showing streak plate of ''E. coli'' DH5α cells containing plasmid pSB1AT3. The pink colouration on the plate is due to the ''rfp'' in the plasmid.
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===Set up culture for minipreps===
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==Setting up of the culture for plasmid miniprep extraction==
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5ml culture with ''E.coli'' from a single colony grown over night.
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Refer to the protocol list for preparation of the overnight culture of ''E. coli'' DH5α cells: [[Team:Newcastle/Growing an overnight cultures| Growing an overnight culture]].
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Tomorrow we will harvest the DNA.
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Tomorrow we will harvest the plasmid DNA.
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==Conclusion==
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We learnt aseptic techniques and some basic wet lab techniques. As you can see in the '''Picture 1''' shown above, our streak plates worked.
{{Team:Newcastle/footer}}
{{Team:Newcastle/footer}}

Latest revision as of 20:07, 21 October 2010

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This is the FIRST DAY of the iGEM lab training for the second group. 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. how to use pipettes.

Contents

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
  • Burnsen burner
  • Conical flasks
  • Orbital shaker

List of techniques

  1. Used electronic balance and made LB broth.
  2. Prepared broth culture
  3. Familiarized with using pipettes of different volumes
  4. Mini-Prep introduction for Tuesday.

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

Tips

  • Use aseptic technique: Treat everything as a pathogen!
  • Work around burnsen burner.
  • Use a control for every experiment.
  • Clean the bench at the end of the day.
  • Heat the flame loop from the middle to the tip till it becomes red hot.
  • Keep plates upside down so that condensation does not damage the colonies.
  • Keep the lids of the plates off for as short time as possible.
  • Loosen all the tops off the vessels before you start.
  • Flame tops of the bottles lightly so as to reduce the chances of contamination.
  • Colony plates should be labelled (name of culture, antibiotics used, our initials and date) at the base and inverted in order to prevent condensation.
  • For individual colony selection, steak across a few times in different directions in order to dilute the number of colonies per unit area.
  • Wear gloves, especially when working with DNA, PCR etc. However, never wear gloves when working near a flame.

Streak plating

Star plate-streaking student, Deena, shows how it is to be done!

Picture 1: Picture showing Star plate-streaking student Deena showing streak plate of E. coli DH5α cells containing plasmid pSB1AT3. The pink colouration on the plate is due to the rfp in the plasmid.

Setting up of the culture for plasmid miniprep extraction

Refer to the protocol list for preparation of the overnight culture of E. coli DH5α cells: Growing an overnight culture.

Tomorrow we will harvest the plasmid DNA.

Conclusion

We learnt aseptic techniques and some basic wet lab techniques. As you can see in the Picture 1 shown above, our streak plates worked.

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