From 2010.igem.org
(Difference between revisions)
|
|
| Line 1: |
Line 1: |
| | {{Team:Newcastle/mainbanner}} | | {{Team:Newcastle/mainbanner}} |
| | | | |
| - | '''25 June 2010'''
| + | =Polymerase Chain Reaction= |
| | | | |
| - | ==Aims== | + | ==Aim== |
| - | The aim of today's Lab practice is to run the products of a PCR reaction on a gel to visualise the inserts and backbones as they separate due to difference in mass.
| + | |
| | | | |
| - | ==Equipment List==
| + | To amplify a desired DNA fragment by using a set of appropriate primers flanking the fragment. |
| | | | |
| - | * Solutions/ enzymes for PCR protocol
| + | ==Materials and Protocol== |
| - | * GoTaq polymerase
| + | |
| - | * Gloves
| + | |
| - | * Agarose gel
| + | |
| - | * Primers(Forward and Results)
| + | |
| - | * dNTPs
| + | |
| - | * Buffers
| + | |
| - | * Ice
| + | |
| - | * Sample DNA
| + | |
| - | * Thermocycler
| + | |
| | | | |
| - | ==Polymerase Chain Reaction protocol==
| + | Please refer to: [[Team:Newcastle/PCR| PCR]] for materials required and protocol. |
| - | | + | |
| - | Polymerase chain reaction (PCR) is a technique used to amplify a single or few a copies of a piece of DNA generating thousands to millions of copies of a particular DNA sequence.
| + | |
| - | # To avoid contamination, wear gloves
| + | |
| - | # To achieve optimum results, always do everything on ice
| + | |
| - | | + | |
| - | A basic PCR set up requires several components and reagents.These components include:
| + | |
| - | #DNA template that contains the DNA region (target) to be amplified.
| + | |
| - | #Two primers that are complementary to the 3' (three prime) ends of each of the sense and anti-sense strand of the DNA target.
| + | |
| - | #Taq polymerase or another DNA polymerase with a temperature optimum at around 70 °C.
| + | |
| - | #Deoxynucleoside triphosphates, the building blocks from which the DNA polymerases synthesizes a new DNA strand.
| + | |
| - | #Buffer solution, providing a suitable chemical environment for optimum activity and stability of the DNA polymerase.
| + | |
| | | | |
| | {| | | {| |
Revision as of 21:05, 21 October 2010
Polymerase Chain Reaction
Aim
To amplify a desired DNA fragment by using a set of appropriate primers flanking the fragment.
Materials and Protocol
Please refer to: PCR for materials required and protocol.
"
