From 2010.igem.org

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.

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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 {{Team:Newcastle/mainbanner}}
-==Chromosomal prep==
+=PCR to test chromosomal prep=
-===Results===
+==Aim==
-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 hundred 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.
+To test whether the chromosome extraction from ''Bacillus subtilis'' ATCC6633 was successful.
-[[Image:Newcastle PCR gel.jpg|400px]]
+==Materials and Procedure==
+Please refer to:[[Team:Newcastle/PCR| PCR]] and [[Team:Newcastle/Gel electrophoresis| Gel electrophoresis]] for materials and protocol.
+We used ''ara'' forward and reverse primers for amplification.
-===Conclusion===
+==Results==
-The aim of this whole experiment was to extract genomic DNA from ''B. subtilis'' strain 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 hundred bps ladder.It worked!
+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.
-==LacI BioBrick Construction==
+[[Image:Newcastle_Genomic_extraction.jpg|400px]]
-===Aims===
+* '''Lane 1''': 100 bp DNA ladder
-* To use PCR to extract ''lacI'' (promoter, ribosome-binding site (RBS) & coding sequence (CDS)) from plasmid pMutin4 and ligate into vector pSB1AT3 in front of red fluorescent protein (RFP).
+* '''Lane 2''': Chromosomal DNA with ara gene 1
+* '''Lane 3''': Chromosomal DNA with ara gene 2
-===Materials===
+==Conclusion==
-* PCR prduct
+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!
-* Restriction enzymes EcoR1 and Spe1
-===Protocol===
-* ''lacI'' PCR product is purified.
-* Purified ''lacI'' is digested with EcoR1 and Spe1.
-===Inference===
-* The purified PCR product is cut to give it sticky ends complementary to the digested pSB1AT3.
-{{Team:Newcastle/footer}}
 {{Team:Newcastle/footer}}

Team:Newcastle/7 July 2010

From 2010.igem.org

Latest revision as of 15:09, 27 October 2010

Contents

PCR to test chromosomal prep

Aim

Materials and Procedure

Results

Conclusion