Assume you have purified the insert-containing plasmid from chloramphenicol or kanamycin-resistant E. coli. You used pBLU...

Assume you have purified the insert-containing plasmid from chloramphenicol or kanamycin-resistant E. coli.

You used pBLU as the vector for the unknown insert project.

You use PCR to identify your unknown insert. What is your positive control, negative control, and experimental group?

Expert Solution

After successful transformation, you need to select colonies on antibiotics, extract and purify the plasmid. For verification of the insert, PCR method can be employed. The experiment will carry following groups:

Negative Control: Negative tube will not have any DNA template means all components required in PCR except insert to rule out any artifact (you supposed to see NO BAND, if you observe then would indicate contamination in your PCR reaction/mixture)

Positive Control: A known DNA template would be there and you supposed to observe the band on agarose gel (BAND SHOW)

Experimental Control: Plasmid extracted from pBLU cells after transformation (carrying desired insert), if its successful, then you supposed to see band at the correct position/size over gel.

gladiator answered 2 years ago

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