Question

In addition to DNA polymerase and primer the polymerase chain reaction also requires

Answer 1

Welcome to OpenStudy.

For PCR you'd need the following:

- A thermal cycler
- Forward and Reverse primers
- T.aq DNA Polimerase
- Deoxynucleotide triphosphates (dNTP)
- Magnesium Chloride (MgCl2)

Answer 2

No, you do not need a thermal cycler, it can be done with hot water and cold water, by hand, though it really sucks to do so. There are many different polymerases, my lab uses Paq not Taq. I believe the best answer is the dNTPs.

Answer 3

It can be done at room temperature (without cycling) if you use "rolling circle" method.
Thomaster just pointed out the components of the most commonly used protocol.

Answer 4

Rolling circle is not polymerase chain reaction. It is a fundamentally different process.

Answer 5

Doing it by hand is not a very realistic solution, it would be hell.
Everyone would use a thermal cycler nowadays.

As for the Taq polymerase, I suppose there could be other DNA polymerases from other thermophiles, it's just they're not being used as often as Taq.
Honestly I've never heard of anyone using a different polymerase.

Answer 6

It was originally done by hand, and there a huge variety of polymerases. I am surprised you have not heard of any others. They are all polymerase though, just things like proof reading and the like.

Answer 7

They both make copies of nucleic acids from a template strand. How is that "fundamentally different"?

They do have different applications though, you wouldn't use the RC method to amplify nucleic acids to run a gel like you would if you were building a diagnostic hybridization-dependent sensor.

Answer 8

The way it is done is fundamentally different. Rolling replication starts at one point, and moves one way, on one strand. PCR makes copies of a targeted region and has a goal of creating only that targeted region, rolling rep does the entire sequence.

Answer 9

lol does the polymerase in PCR not start at the primer site (one point) and move in a 5' to 3' direction copying the sequence downstream?

This is exactly what happens in both cases.

In RC you can isolate the region you want to copy, it doesn't need to be an intact whole strand.

The main difference is that in PCR the copies are fragments, and in RC the copies are 1 long continuous strand (concatamer).

ps. Sure the reaction conditions are not the same, but that was my point, you can perform amplification of nucleic acids at room temp using the RC method.

Answer 10

No, you need to read up on it more. It pulls off a single region and requires the that the gDNA is cut first. It has PCR requires no cut, and the goal of PCR is not the same as the goal of rolling rep. Just because the both replicate DNA it doesn't follow they are the same thing.

Answer 11

@aaronq the question was about polymerase chain reaction.