Lab 10: Isolation of the plasmid, and endonuclease restriction excision of the DNA insert
Lab Report 10 Plasmid Isolation and Restrictions Enzyme
DNA Plasmid Isolation/Restriction Digestion Laboratory Report
Write down the following for your miniprep solution:
A260 reading __0.116______
A280 reading _____0.058___
A260/A280 ratio __Need to figure out and show work here_________
Also report here whether the A260/A280 ratio you reported above is a good DNA ratio. (Google what is a good DNA ratio).
Based upon your measured A260/A280 ratio what can you say about the quality of your DNA plasmid solution?
Look up a DNA phenolic extraction/ethanol precipitation protocol online (Googling it should bring up several lab protocols other people have put online. Describe the steps for such a protocol below, and explain for each step what is happening to the cells (what you are isolating the DNA from) and/or DNA (what you are trying to isolate).
What kind of nucleases are BamHI and EcoRI? Be specific.
What were the original sources of BamHI and EcoRI?
Why endonucleases work best at 37 °C?
Describe exactly where BamHI and EcoRI cut the DNA (i.e. the exact sequences). For this nuclease show exactly what the DNA cut ends would look like (i.e. the exact sequences). Are the ends sticky or blunt? Why?
