Difference between revisions of "Week Four"
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− | ===June 8th=== | + | ===June 8th- 17th=== |
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+ | '''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: | |
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− | ''' | + | '''[[Step 1 - Taking dry DNA from wells]]''' |
− | + | '''[[Step 2 - Transforming competent cells]]''' | |
− | + | '''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. | |
− | + | '''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]]''' | |
− | ''' | + | '''Step 7 - [[Gel Electrophoresis]]''' |
− | + | '''Step 8- [[Ligation]]''' | |
− | + | '''Step 9- [[Transformation and Inoculation]]''' | |
− | + | ===June 9th=== | |
− | + | [[The June 9th Image Gallery]] | |
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− | + | Images of the various results attained: | |
− | + | <gallery> | |
+ | File:Gelimage090609.png|Agarose gel electrophoresis image of digested K112808 | ||
+ | File:Not working.jpg|Failure 1 | ||
+ | File:PBAD-insert f.png| The pBAD Insert | ||
+ | File:Failed Ligation.png| The Failed Ligation Attempt | ||
+ | </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
Images of the various results attained: