00:01
For one, you want to clone a fragment of dna into a plasmid vector such as pbr322.
00:06
The plasmid is 4 .36 kv long and has a single restriction site for the restriction and then bamh1.
00:13
Now, if you digest the pbr322 with bamh1, how many linear fragments would you have? so this is pbr322.
00:27
It has a single bamh1 site.
00:37
So let's say it's here.
00:38
So once you cut the plasmid with the bamh1, the enzyme is going to recognize the bamh1 site and cut it.
00:50
So it only is going to cut once right here.
00:54
So once you cut the circular plasmid once with bamh1 enzyme, it is going to linearize.
01:04
And the size of this fragment is going to be 4 .36 kv.
01:10
And you only produce one fragment.
01:13
So the correct answer is going to be a, one.
01:16
And this fragment is 4 .36 kv long.
01:21
So basically you cut a circle open so it become linearized.
01:26
So this is the first question.
01:28
So i draw the figure to show you what it looks like.
01:31
The second question.
01:33
Now, you have pbr322 has a single bamh1 site and a single bspmi site.
01:41
A double digestion produced two fragment of approximately 690 base pairs, 3670 base pair.
01:49
Map the two sites on a circular plasmid.
01:53
If the bamh1 is at the beginning of the plasmid, tetracycline resistant gene, and a gene is about 1200 base pair long, is the bamh1, bspmi site also in the gene? so let's say you have a plasmid circular...