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the gel is numbered from left to right, the first clone from left is ladder, the

ID: 81545 • Letter: T

Question

the gel is numbered from left to right, the first clone from left is ladder, the second is 1RS, and so on...

clone 4R, 5R, and 6R contains one type of DNA. 7R, 8R, and 9R contains another sample of DNA.

Summarize your conclusions regarding your plasmids using the table below: whether they have an insert or not, and which one(s) you would predict have the correct LacZa-GFP fusion you tried to obtain in this lab!

If some of your clones have an unexpected restriction pattern, do mention it and explain, based on your gel result and looking at either or both digests, why you think the plasmid has an unexpected structure (it does happen sometimes!).

                        Insert?                        Orientation?                           Unexpected pattern?

                        Y/N                 Correct/incorrect                 Y/N (if Y, explain why at the                                                                                                                                                            bottom of the page)

Clone 1:

Clone 2:

11 21 3]4]5]6]71 8 Blo-Rad Laboratories CM LL016 8 L 9 S

Explanation / Answer

1) so that we can check whether and how many Bam H1 site is present in the vector. if our plasmid has an insert then after single digestion either with Bam H1 or AlwN1 we will get upshift in DNA band as compare to vector control digested with the same restriction enzyme.

2) for clone no 1 digested with either with Bam H1 or AlwN1 or with both the enzymes, we got the same band size as compare to vector control digested with the same restriction enzyme. means.it has no insert.

3) for clone no 2 digested with either with Bam H1 or AlwN1, we got 2 bands, suggesting that our insert also contains restriction sites for both the enzymes resulting two visible bands in the gel. so we got the unexpected pattern.

4) for clone no 2 digested with Bam H1 and AlwN1, we got 3 bands, suggesting that our insert also contains restriction sites for both the enzymes resulting three visible bands in the gel. so we got the unexpected pattern.