How were restriction enzymes used in pGLO?

Restriction digests also form part of the foundation for molecular biology in general. Recombinant DNA molecules (such as the pGLO plasmid) are traditionally made by cutting various desired DNA fragments with restriction enzymes, then ligating (joining) the pieces together.

What is a restriction map What is a genetic map?

Genetic mapping or linkage mapping can be used to indicate the relative order of genes on a chromosome. A restriction map is another type of DNA map that roughly describes the relative positions of genes by breaking apart sections of DNA at locations known as restriction sites.

How do you do restriction mapping?

One common method for constructing a restriction map involves digesting the unknown DNA sample in three ways. Here, two portions of the DNA sample are individually digested with different restriction enzymes, and a third portion of the DNA sample is double-digested with both restriction enzymes at the same time.

How do you find the number of restriction fragments?

First, work out the frequency of occurrence of the restriction site as 1-in-x bases, as explained in the example for the Intermediate level calculation. Then take the size of the DNA in kb (kilobases) and multiply by 1000 to get the size in bases. Divide this by x and round to the nearest whole number.

What are the restriction sites of the pGLO plasmid?

Multiple cloning site — a region containing restriction sites (NdeI, HindIII, EcoRI, etc.), sequences that permit the insertion or deletion of a gene of interest Bacteria transformed with the pGLO plasmid are selected by ampicillin resistance and when induced to express GFP, they glow fluorescent green under UV light!

What are the 3 restriction enzymes in pGLO?

The restriction enzymes are: The first three (Hind III, Pvu I, and Ssp I) are 6-base cutters; their restriction sites are 6 nucleotides long. Note that Hind III and Pvu I leave sticky ends, while Ssp I leaves blunt ends: both strands are cut at the same place. Sat I is different from the others.

How to see the pGLO sequence in snapgene?

Bacterial transformation plasmid for regulated expression of GFP. To see this sequence with restriction sites, features, and translations, please download SnapGene or the free SnapGene Viewer.

How is the pGLO plasmid modified for jellyfish?

•The wild-type jellyfish gene has been modified by a biotech company; specific mutations were introduced into the DNA sequence, which greatly enhance fluorescence of the protein. •This modified form of the GFP gene has been inserted into our pGLO plasmid. GFP