Difference between revisions of "Week Four"

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(June 8th- 17th)
(June 9th)
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File:Gelimage090609.png|Agarose gel electrophoresis image of digested K112808
File:Gelimage090609.png|Agarose gel electrophoresis image of digested K112808
File:Not working.jpg|Failure 1
File:Not working.jpg|Failure 1
File:PBAD-insert f.png| the PBAD Insert
File:PBAD-insert f.png| The pBAD Insert

Revision as of 08:45, 24 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:

For June 15th:

These bands will be kept overnight in 37 degrees Celsius.

Then these will be run through a preparative gel which takes about 3 hours.

The next step is to elute the released products which wold take 1.5 hours.

CIP treatment will be done [Calf Intestine Phosphate] This treatment is given to only the vector of Lysis (Process will take 1.5 hour) This is done so that the vector doesn't self ligate.

This enzyme will then be inactivated at 65 degree C.

This vector needs to be purified using column or chloroform