During cloning H If the phosphatase didnt work explain how t

During cloning,   

H) If the phosphatase didn’t work, explain how this would affect the relative number of blue and white colonies.

     I) If the restriction enzyme only digested half of the circular vector molecules to linear molecules, explain how would this affect the relative number of blue and white colonies.

     J) Explain how you could get pale blue colonies?

In colony PCR

1. Explain/discuss each of the three \"steps\" for PCR.

Solution

1) the three steps are

Denaturation - in this step the DNA is heated to 94\'C to separate the two DNA strands..!!

Aneeling - in this step the primer is being aneled to the template at 54\'C..!!

Extension - then the DNA polymerase enzyme extend the DNA primer result in the formation of the new strand ar 72\'C..!!

J) the blue color colonies is formed in BLUE/WHITE RECOMBINATION screening whre we can differentiate between the two bacteria which had been recombinant or not..!! The blue colour is due to the breakdown of the xgal protein.. It follows a mechanisms called alpha complentation... The bacteria has lac - Z gene which can form omega portion Wheraes the Plasmid has the omega portion of gene.. When the given Plasmid gets transformed into a bacterial cells the Plasmid forms the alpha portion combined with omega form lac-Z which breakdown xgal substrate and gives blue color..!!