Predict the results from your PCR amplification of GFP by ex

Predict the results from your PCR amplification of GFP by explaining what you would see on the agarose gel.

You attempted to amplify a portion of GFP (green fluorescent protein) gene from three different DNA samples:
1. E. coli wild type grown in LB
2. E. coli + pGLO grown in LB + amp
3. E. coli + pGLO grown in LB + amp + arabinose

Be sure to include a DNA ladder of known DNA sizes.

The GFP PCR product is 714 bp.

Solution

The PCR reaction aims at amplyfying the \'GFP gene sequence\' and not the \'GFP Gene product\', which in this case is Green Fluorescent Protein (GFP) itself. As such, the following samples will have the gene sequence for GFP

A) E. coli + pGLO grown in LB + amp
B) E. coli + pGLO grown in LB + amp + arabinose

Both the above samples contain the pGLO vector, which contains the cloned GFP gene. Hence, PCR products from both these samples will show a band in the agarose gel, which corresponds to a bp of 714.

Sample 1 will not have any band, since it does not contain the GFP-containing pGLO vector.