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You wish to perform a PCR and the target sequence to which one of your primers will bind is:5'-CACGAGTCAGGGTCCTT-3'You would like to add a restriction sit...

Question

You wish to perform a PCR and the target sequence to which one of your primers will bind is:5'-CACGAGTCAGGGTCCTT-3'You would like to add a restriction site to the primer that will anneal to this site. This will be the recognition site for the enzyme BamH1 which is 5'-GGATCC. Design the primer that will anneal to the sequence above and incorporate the site for BamH1:

You wish to perform a PCR and the target sequence to which one of your primers will bind is: 5'-CACGAGTCAGGGTCCTT-3' You would like to add a restriction site to the primer that will anneal to this site. This will be the recognition site for the enzyme BamH1 which is 5'-GGATCC. Design the primer that will anneal to the sequence above and incorporate the site for BamH1:



Answers

Designing primers II. You wish to amplify a segment of DNA from a plasmid template by PCR with the use of the following primers: $5^{\prime}-$ GGATCGATGCTCGCGA-3' and $5^{\prime}-$ AGGATCGGGTCGCGAG-3'. Despite repeated attempts, you fail to observe a PCR product of the expected length after electrophoresis on an agarose gel. Instead, you observe a bright smear on the gel with an approximate length of 25 to 30 base pairs. Explain these results.

Counsel for the question here. Primer. We have to design to a primer here. Primer one five c c t c g A g d. See a BC G 80 d C t g three and we HAVE Raimond to fly five c g c g C A C A D C e g A gsc g A C C A three

Yes, we need to the size. What's Primer Wigan's using after this gene? And we've been given the sequence sequence. It is person three primes it like from. So our confidence is going to be five friends, three friends. So we just need to make sure we note that down. Isn't that sweet? 3.5 from ends. And we're just going to write down V compliments, which is the unites we're going seven as a teen, uh, finding instead of a year or so they just just writing down be, uh, the compliments has getting check. It's on. Yeah, there we go. Is three strands compliments to what we've been given I'm notes again. Bar fronts of reprint to complement the free fun Seeing five for instructions and given

And so for the question here, right, These sequences of the two 12 residues primary state could be used to amplify the loving DNA segments by PCR. So here, the Cuban segment we have here first look at the first of all types and the complementary basis of the last. Well, nobody's do know these segments of the primaries to be used. So here we have Eddie Mm G g c A D A G see and C d g a c c e g c g g c g c c. So look at the 1st 12 nucleotides and the complementary basis of the last 12 nucleotides to know the sequences of the primaries to be used.

Hi, everyone. My name is Eric and let's review problem eight. So, after reading the question, you should find that there are two things that this question is asking for. The first part is taking the amino acid sequence and translating it back into a nucleotide sequence. The second question is asking about the middle searing and what happens when it changes to assisting. What would that sequence look like? So let's focus on the first part. So after looking at the amino acid sequence, you should find that there are 17 amino acids, and that means that there is going to be 51 nucleotides. That is because each amino acid is comprised of three nucleotides. So we call that a code on and let's write out the amino acid sequence here. So first they have a sparrow Jing searing glide scene Dining history Dean Pro lean glycerine Life scene, Loose scene, Alan. I mean searing trip to fend phenylalanine. I'm running out of space here. We'll just continue at the bottom here, so just know that this is gonna be a continuation. So then we have very lean Glad I seen again a spar Janine and finally searing So this will be the beginning with the estrogen in up here and the the ending will be the Syrian here. So in order to translate this amino acid sequence two nucleotides, we have to look at what's called DNA code on chart or table, and I recommend that you use this to translate each amino acid to its nucleotide sequences. So each of these amino acids are going to be comprised of three nucleotides each. And when you look at the table, there's going to be differences in the sequences. But most of the time, if you pick one of the amino acids, that should be the sequence that corresponds to that amino acid. So, for example, I'll be solving this part of the this solved the problem here. So let's start with a Spar Jenning. So far, Johnny, there are different combinations of nucleotides, but I picked one so the first one will be a T and then searing TCG glassine G T and the dining A. T g. This state Dean C. A c pro lean CCG glycerine G a. Listen a g loosen t t a. Allan e g c c searing T c T trip to fan T g. And finally nein T t c veiling g t a glazing g a spar Jinyan again a 80 and finally searing T c A. So this is a perfectly valid nucleotide sequence that corresponds to the amino acid sequence, right? The differences. The nucleotides is DNA and amino acids are for protein, so that would be code for protein. And that is the first part of the question where they're asking you to translate it from amino acid back to nuclear side. And so the second part is talking about searing the middle searing, to be exact. So that would be this one right here. And they want to change this searing into assisting. So by using the DNA code on table, you would be able to change the searing code to assisting code. And how would we do that? T. C t, in my example here, can be changed to Sistine by changing the middle seat from T C T to T g t. So this would be a way to change it from searing to Sistine. And this is a perfectly valid example. And there are many other ways that you can do this there are different combinations as long as they match. The amino acid is perfectly valid, so I hope this helps bring a little bit of clarification into understanding how to change from amino acid to nucleotide sequence. A key component will be using this table. It's a really important tool and also hope you that you understand how to change the Syrian to assisting and how the sequence is related to the amino acid. Thanks for listening and see you next time.


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