After we have cut and dissolve our DNA from the gel, we can then add ethonal so the DNA falls out and we also add special beads to bind to the DNA so it can separate and stay at the bottom of the test tube. After that we can dissolve the DNA in free water. And we have now successfully extracted our DNA!
DNA can be added to a plasmid or a topovector, which will allow us to insert our DNA into bacteria during the process of transformaiton. This procedure is also called ligation.
Transforamtion:
Transformation is used to "get more DNA." When the plasmid is added to the bacteria, the bacteria grows and the plasmid, containing our targeted DNA, will grow with it.
1) thaw the bacteria
2) add ligation to bacteria
3) incubate ligation and bacteria on ice for 30 min.
4) heat shock > at a higher temp. the bacteria will expand to release heat. And when it is expanded, the plasmid has the ability to travel inside the bacteria. Then the temp. decreases again and the bacteria closes, capturing the plasmid inside.
5) plate the bacteria > the bacteria is given time to grow, and the bacteria with antibiotic resistance will be the only ones left living for our use.
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| plating the bacteria |
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| bacteria after growth |
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Next week we are looking forward to getting the sequencing results, which will give us the neucleotide base sequence of our gene of interest, answering the first part of my project...
Is gene 5 a real gene?
