Let's suppose you want to clone a gene that has never been...

Let's suppose you want to clone a gene that has never been analyzed before by DNA sequencing. Which of the following statements do you agree with the most? a. Do PCR to clone the gene because it is much faster. b. Do PCR to clone the gene because it is very specific and gives a high yield. c. You can't do PCR because you can't make forward and reverse primers. d. Do cloning using a vector because it will give you a higher yield. e. Do cloning by insertion into a vector because it is easier than PCR.

Let's suppose you want to clone a gene that has never been analyzed before by DNA sequencing. Which of the following statements do you agree with the most?
a. Do PCR to clone the gene because it is much faster.
b. Do PCR to clone the gene because it is very specific and gives a high yield.
c. You can't do PCR because you can't make forward and reverse primers.
d. Do cloning using a vector because it will give you a higher yield.
e. Do cloning by insertion into a vector because it is easier than PCR.

Key Concepts

Gene Cloning

Gene cloning is the process of making multiple copies of a gene or DNA sequence by inserting it into a vector, such as a plasmid, which is then introduced into a host organism. This method can be used even when the gene's sequence is not fully known, because it does not require prior sequence information for amplification.

Polymerase Chain Reaction (PCR)

PCR is a widely used technique to amplify specific segments of DNA. It requires the design of short sequences, called primers, that flank the target region. The success of PCR depends on knowing parts of the gene sequence to create these primers, making it less suitable for cloning a gene that has never been sequenced.

Primer Design

Primer design involves creating short, single-stranded DNA sequences that are complementary to the start and end of the target DNA segment. Effective primer design is critical for PCR, as it ensures the specific amplification of the desired sequence. Without prior sequence information, designing effective primers is not feasible.

Transcript

00:01 Now in this question, we are addressing the differences between cloning by insertion into a vector versus cloning by pcr.

00:50 Okay, so now we got to see what the difference is between these two.

00:56 Well, first of all, cloning by insertion into a vector is a nice advantage because it is very accurate.

01:06 You're using very complex methods here.

01:11 Very accurate is probably the wrong word because it's not that specific, but it's, i would say, very likely to succeed.

01:29 So let's call that very successful, although, you know, it's going to have a couple of problems, which we'll address now.

01:39 Even though it's very successful on cloning even like unknown genes, in other words, you have a gene, but you don't really know the sequence, you can do very well by cloning insertion into a vector.

02:05 It is a kind of genetic engineering.

02:10 So it is very complex to do.

02:18 You're essentially inserting a gene into another organism.

02:23 It's genetic engineering.

02:34 Also, not all cells will have the clone.

02:46 So you have to differentiate.

02:53 You know, did you have that clone? and it was like, why? why? if you have this, why isn't it cloned all the time? again, because of the way we do genetic engineering.

03:07 When we do genetic engineering, we insert the gene into the vector.

03:13 But then the bacteria can take it up by transformation.

03:20 So bacterial transformation is not 100%.

03:35 So we have to take the time to identify what colonies have the clone of interest that we want.

03:44 Okay? and then you're saying, well, if that's, you know, so complicated, why do it? again, because it is very successful at cloning an unknown gene.

03:56 You just have to go through a lot of hoops.

03:59 So it is kind of slow, but worth it if you want to clone a gene.

04:09 Okay, so it's very complex, low, so that gives you a low yield at first.

04:21 Once you isolate the bacteria that have the gene you clone, and then you're in business.

04:27 Okay, because then you only culture those bacteria and off you go.

04:32 However, we're talking here about a gene that's never been analyzed before, and we want to clone more of them so we can study it better.

04:42 Well, pcr is very tempting here because pcr polymerase chain reaction is claim to fame...