Team:Newcastle/25 June 2010

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{{Team:Newcastle/mainbanner}}
{{Team:Newcastle/mainbanner}}
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'''Friday'''
 
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===Aims===
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=Polymerase Chain Reaction=
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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.
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===Equipment List===
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==Aim==
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* Solutions/ enzymes for PCR protocol
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To amplify a desired DNA fragment by using a set of appropriate primers flanking the fragment.
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* GoTaq polymerase
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* Gloves
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* Agarose gel
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* Primers(Forward and Results)
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* dNTPs
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* Buffers
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* Ice
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* Sample DNA  
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* Thermocycler
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===Polymerase Chain Reaction protocol===
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==Materials and Protocol==
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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.
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Please refer to: [[Team:Newcastle/PCR| PCR]] for materials required and protocol.
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# To avoid contamination, wear gloves
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# To achieve optimum results, always do everything on ice
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A basic PCR set up requires several components and reagents.These components include:
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==Result==
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#DNA template that contains the DNA region (target) to be amplified.
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#Two primers that are complementary to the 3' (three prime) ends of each of the sense and anti-sense strand of the DNA target.
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#Taq polymerase or another DNA polymerase with a temperature optimum at around 70 °C.
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#Deoxynucleoside triphosphates, the building blocks from which the DNA polymerases synthesizes a new DNA strand.
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#Buffer solution, providing a suitable chemical environment for optimum activity and stability of the DNA polymerase.
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{|
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[[Image:8062010.jpg|400px|center]]
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|[[Image:fri-gel.png|300px|thumb]]
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|[[Image:Newcastle gel 20100625.jpg|300px|thumb]]
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'''Image 1''': Image of the gel showing PCR products.  
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|}
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* '''Lane 1''': 1kb DNA ladder
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* '''Lane 2''': Sample 1
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* '''Lane 3''': Sample 2
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* '''Lane 4''': Sample 3
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* '''Lane 5''': Sample 4
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* '''Lane 6''': Sample 5
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* '''Lane 7''': Sample 6
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* '''Lane 8''': Sample 7
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* '''Lane 9''': Sample 8
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* '''Lane 10''': Sample 9
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* '''Lane 11''': Sample 10
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* '''Lane 12''': Sample 11
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* '''Lane 13''': Sample 12
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* '''Lane 14''': Sample 13
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* '''Lane 15''': Sample 14
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{{Team:Newcastle/footer}}

Latest revision as of 13:54, 27 October 2010

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Contents

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.

Result

8062010.jpg

Image 1: Image of the gel showing PCR products.

  • Lane 1: 1kb DNA ladder
  • Lane 2: Sample 1
  • Lane 3: Sample 2
  • Lane 4: Sample 3
  • Lane 5: Sample 4
  • Lane 6: Sample 5
  • Lane 7: Sample 6
  • Lane 8: Sample 7
  • Lane 9: Sample 8
  • Lane 10: Sample 9
  • Lane 11: Sample 10
  • Lane 12: Sample 11
  • Lane 13: Sample 12
  • Lane 14: Sample 13
  • Lane 15: Sample 14


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