Team:Newcastle/7 July 2010

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(Chromosomal prep)
 
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=Chromosomal prep=
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=PCR to test chromosomal prep=
==Aim==
==Aim==
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==Materials and Procedure==
==Materials and Procedure==
Please refer to:[[Team:Newcastle/PCR| PCR]] and [[Team:Newcastle/Gel electrophoresis| Gel electrophoresis]] for materials and protocol.
Please refer to:[[Team:Newcastle/PCR| PCR]] and [[Team:Newcastle/Gel electrophoresis| Gel electrophoresis]] for materials and protocol.
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We used ''ara'' and ''sac'' forward and reverse primers for amplification.
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We used ''ara'' forward and reverse primers for amplification.
==Results==
==Results==
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We used Gel Electrophoresis to test whether we have the DNA we wanted. As we already know, the ''ara'' gene is about 300bps. We used 100 bp DNA ladder as a guidance for our bands. The two bands produced were from two separate ''ara'' cultures and they indeed show the bands in the same region, i.e. 300bps.  
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We used gel electrophoresis to confirm the quality of our DNA. As we already know, the region our primers amplify is about 300bps. We used 100 bp DNA ladder as a guidance for our bands. The two bands produced were from two separate chromosomal preps.  
[[Image:Newcastle_Genomic_extraction.jpg|400px]]
[[Image:Newcastle_Genomic_extraction.jpg|400px]]

Latest revision as of 15:09, 27 October 2010

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Contents

PCR to test chromosomal prep

Aim

To test whether the chromosome extraction from Bacillus subtilis ATCC6633 was successful.

Materials and Procedure

Please refer to: PCR and Gel electrophoresis for materials and protocol. We used ara forward and reverse primers for amplification.

Results

We used gel electrophoresis to confirm the quality of our DNA. As we already know, the region our primers amplify is about 300bps. We used 100 bp DNA ladder as a guidance for our bands. The two bands produced were from two separate chromosomal preps.

Newcastle Genomic extraction.jpg

  • Lane 1: 100 bp DNA ladder
  • Lane 2: Chromosomal DNA with ara gene 1
  • Lane 3: Chromosomal DNA with ara gene 2

Conclusion

The aim of this whole experiment was to extract genomic DNA from Bacillus subtilis ATCC 6633. In order to test whether we had extracted the DNA, we first used PCR to amplify and then used Gel Electrophoresis to compare the bands of ara genes to the 100 bp ladder. It worked!


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