Difference between revisions of "Week Four"

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(Created page with '===June 8th=== '''''A Brief Description of What We Will Do''''' The Plasmid preparation process | [[The Pla...')
 
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===June 8th===
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===June 8th- 17th===
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'''''[[A Brief Description of What We Will Do]]'''''
 
  
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[[Preparing the pBAD and Lysis plasmids for gel electrophoresis|The Plasmid preparation process]] | [[The Plasmid Preparation Miniprep Gallery]]
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'''What are we trying to do?'''
  
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We want the bacteria to lyse, to kill itself from the inside.
  
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''' A Brief of what we did: ''' <br/>
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Well then, to do that we had to get a lysis gene in it. The thing is, with the lysis gene needs to be turned on by this specific substance called a '''promoter'''
  
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The plasmids that contained the pBAD and Lysis genes that were prepared were extracted from the bacteria cultures. The unwanted protein around the DNA was then removed. After that the DNA was cleansed of any remaining salt. The plasmids were then cut at certain points such that only the required gene could be extracted. These cut parts were prepared for gel electrophoresis.
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There are two kinds of promoters, ones that make the gene they're attached to , be 'on'  all the time.
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Then, you have ones that turn on when you want them to, in our case, when you add a mystery enzyme.
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'''Our Step-wise Process:'''
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How we tried to do this:
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'''[[Step 1 - Taking dry DNA from wells]]'''
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'''[[Step 2 - Transforming competent cells]]'''
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'''Step 3 - Picking a single colony.'''
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'''Step 4 - Inoculating broth with Ampicillin-R''' (an antibiotic) and letting it grow for 18 hours.
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'''Step 5 - Using the resulting culture for [[mini-prep.]]''' - ''a process used to purify plasmids and yields clean, usable DNA.''
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'''Step 6 - [[Digesting the DNA]]'''
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'''Step 7 - [[Gel Electrophoresis]]'''
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'''Step 8-  [[Ligation]]'''
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'''Step 9-  [[Transformation and Inoculation]]'''
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===June 9th===
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[[The June 9th Image Gallery]]
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Images of the various results attained:
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<gallery>
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File:Gelimage090609.png|Agarose gel electrophoresis image of digested K112808
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File:Not working.jpg|Failure 1
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File:PBAD-insert f.png| The pBAD Insert
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File:Failed Ligation.png| The Failed Ligation Attempt
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</gallery>

Latest revision as of 06:56, 26 June 2009

June 8th- 17th

What are we trying to do?

We want the bacteria to lyse, to kill itself from the inside.

Well then, to do that we had to get a lysis gene in it. The thing is, with the lysis gene needs to be turned on by this specific substance called a promoter

There are two kinds of promoters, ones that make the gene they're attached to , be 'on' all the time. Then, you have ones that turn on when you want them to, in our case, when you add a mystery enzyme.

Our Step-wise Process:

How we tried to do this:

Step 1 - Taking dry DNA from wells

Step 2 - Transforming competent cells

Step 3 - Picking a single colony.

Step 4 - Inoculating broth with Ampicillin-R (an antibiotic) and letting it grow for 18 hours.

Step 5 - Using the resulting culture for mini-prep. - a process used to purify plasmids and yields clean, usable DNA.

Step 6 - Digesting the DNA

Step 7 - Gel Electrophoresis

Step 8- Ligation

Step 9- Transformation and Inoculation

June 9th

The June 9th Image Gallery

Images of the various results attained:

  • Agarose gel electrophoresis image of digested K112808

  • Failure 1

  • The pBAD Insert

  • The Failed Ligation Attempt

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