00:01
So in this case, you have a scenario.
00:02
It doesn't really say about a question, but i think i can explain to you why you've seen 13 white colonies and 80 blue colonies.
00:12
So you start out with a plasmid.
00:15
It's a adaptive plasmid with smr, antibiotics -resistant gene, and it has a lacz gene, which has multiple cloning site, which has a lot of restriction enzyme site.
00:31
Then you also have a yfp.
00:34
You want to clone that into the vector.
00:37
So the very first step we call this restriction digest.
00:47
This will be a step that you cut both dna fragments with the same restriction enzyme.
00:54
So i can't see really the graph, so i don't know which enzyme will be used.
01:00
But they have to be the same restriction enzyme so that yfp can be cloned into the plasmid.
01:09
Once the restriction enzyme digestion is done, you're going to do this reaction we call ligation.
01:16
So the two fragments with the same restriction site on the side can be cloned together.
01:24
So you you end up having a recombinant plasmid that has the smr antibiotic resistant gene, which will make it survive on the spectromycin medium.
01:38
And due to the insertion, you can see that the lacz gene is disrupted by yfp because it inserts inside of the sequence, so lacz disrupted.
01:51
Now, the next step, you introduce the plasmid with the yfp insertion into the bacteria by transformation.
02:18
So there are several scenarios.
02:20
The first scenario, this is a bacteria, and it has its chromosome dna, but it doesn't have a plasmid...