You are cloning a gene into the vector, pUC19 (Figure 1). You are inserting your gene into the
UNIQUE BamHI site in the multiple cloning site found in the lacZ gene (Figure 1 and Figure 2).
After you digest pUC19 with BamHI, you mix the digested plasmid DNA with your gene (with
BamHI sticky ends), and DNA ligase. You then transform E. coli cells with the ligation mixture,
plate the cells on the selective LB media, with ampicillin and X gal, and incubate the plate
overnight at 37C. The next day, you observe your transformation plates. An example plate is
shown in Figure 3. Figure 3: Bacteria transformed with pucis ligation mixtureFigure 3: Bacteria
translormed with pucis ligation mbxture LB media agar plate with ampiollin and X-gal (5-8romo-4
Chloro-3-Indoly 1 --0-Galactopyranoside) PROMPT: Examine the transformation plate shown in
Figure 3. Your goal is to identify the a colony containing at recombinant plasmid DNA (pUC19 with
your gene). Please answer these two questions: 1. Will you select a "white" or a "blue" colony? 2.
Why are you selecting that colony?.
You are cloning a gene into the vector pUC19 Figure 1 Yo.pdf
1. You are cloning a gene into the vector, pUC19 (Figure 1). You are inserting your gene into the
UNIQUE BamHI site in the multiple cloning site found in the lacZ gene (Figure 1 and Figure 2).
After you digest pUC19 with BamHI, you mix the digested plasmid DNA with your gene (with
BamHI sticky ends), and DNA ligase. You then transform E. coli cells with the ligation mixture,
plate the cells on the selective LB media, with ampicillin and X gal, and incubate the plate
overnight at 37C. The next day, you observe your transformation plates. An example plate is
shown in Figure 3. Figure 3: Bacteria transformed with pucis ligation mixtureFigure 3: Bacteria
translormed with pucis ligation mbxture LB media agar plate with ampiollin and X-gal (5-8romo-4
Chloro-3-Indoly 1 --0-Galactopyranoside) PROMPT: Examine the transformation plate shown in
Figure 3. Your goal is to identify the a colony containing at recombinant plasmid DNA (pUC19 with
your gene). Please answer these two questions: 1. Will you select a "white" or a "blue" colony? 2.
Why are you selecting that colony?
