b. Generate three ligation reactions with 1:1, 2:1 and 3:1 molar ratios of the insert and...

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Biology

b. Generate three ligation reactions with 1:1, 2:1 and 3:1 molarratios of the insert and the vector DNA. Your ligation shouldinclude all components in a 30 ÂµL reaction. Keep the vector amountfixed at 60 ng per ligation reaction. You are provided with 10 xligase buffer and DNA ligase (0.5 U/ÂµL) to set up yourligations.

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In princile cloning in any vector is done by cleaving ofcircular plasmid DNA with one or more restriction enzymes and thenligated to foreign DNA of interest with compatible terminals Thisrecombinant plasmid is then transformed into an appropriate strainof EColi Screening of resultant transformed colonies are done byusing polymerase chane reaction PCR method digestion withrestriction enzyme or by hybridization experiments in order toidentify the desired DNA sequenceCLONING DNA FRAGMENT WITH PROTRUDING ENDS5 or 3 end with 16 basepair in length of protruding end iscreated by digesting the See Answer

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