Latest revision as of 22:12, 27 October 2010

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Problem

Genomic DNA extraction experiment

Aims

The aim of today's experiment is to extract genomic DNA from both B. subtilis strains 168 and 3610. The genes necessary for the swarming BioBrick and rocF BioBrick will then hopefully be obtained from the genomic DNA using PCR.

Protocol

Please refer to: DNA extraction of B. subtilis for materials required and protocol.

Discussion

At the end of the DNA precipitation step, we observed a small white pellet in all the eppendorf tubes.

Conclusion

The experiment was a success! The quality of the extracted DNA will be checked by using PCR on 28th July, 2010.

Preparation for cloning of the rocF BioBrick

Results

Yesterday, we transformed E. coli DH5α with pSB1C3 and [http://partsregistry.org/Part:pSB1AK3 pSB1AK3] with [http://partsregistry.org/Part:BBa_B0014 BBa_B0014]. After checking the plates today for colonies we observed lots of pink colonies on the pSB1C3 plates and numerous white colonies on the [http://partsregistry.org/Part:pSB1AK3 pSB1AK3] with [http://partsregistry.org/Part:BBa_B0014 BBa_B0014] plates. Plates were then stored at 4°C as colonies will be used for minipreps.

@@ Line 1: / Line 1: @@
 {{Team:Newcastle/mainbanner}}
-==Genomic DNA extraction experiment==
+=Genomic DNA extraction experiment=
 [[Image:Newcastle alan chromosome.jpg|thumb|200px|right]]
 [[Image:Newcastle ice chromosome.jpg|thumb|200px|right]]
-===Aims===
+==Aims==
-The aim of today's experiment is to extract genomic DNA from both ''B. subtilis'' strains 168 and 3610. The genes necessary for the swarming biobrick and ''rocF'' biobrick will then hopefully be obtained from the genomic DNA using...
+The aim of today's experiment is to extract genomic DNA from both ''B. subtilis'' strains 168 and 3610. The genes necessary for the [[Team:Newcastle/Swarming|swarming BioBrick]] and [[Team:Newcastle/Urease|''rocF'' BioBrick]] will then hopefully be obtained from the genomic DNA using PCR.
-===Protocol===
-* Please refer to the [[Team:Newcastle/DNA extraction| DNA extraction of ''B. subtilis'']] page
-===Discussion===
-At the end of the DNA precipitation step, we did observe a small white pellet in all the eppendorf tubes.
-===Conclusion===
-The experiment was a success! The content and purity of the extracted DNA will be checked by using PCR on 28th July, 2010.
-==Gel Electrophoresis==
-[[Image:P7270470.JPG|thumb|300px|right]]
-===Protocol===
-* Please refer to the [[Team:Newcastle/Gel electrophoresis|gel electrophoresis]] page for full protocol.
-===Discussion===
-* Unfortunately no bands were observed on the gel.
+==Protocol==
+* Please refer to: [[Team:Newcastle/DNA extraction| DNA extraction of ''B. subtilis'']] for materials required and protocol.
-===Conclusion===
+==Discussion==
+At the end of the DNA precipitation step, we observed a small white pellet in all the eppendorf tubes.
-As no bands were observed on the gel, the PCR did not work. This could be due to a number of reasons: It could be due to the DNA not being extracted or that we might have used the wrong primers. It could also be caused by the small size of the fragment that we amplified which ran off the gel because we ran the gel for too long.
+==Conclusion==
+The experiment was a success! The quality of the extracted DNA will be checked by using PCR on 28th July, 2010.
+=Preparation for cloning of the rocF BioBrick=
+==Results==
+Yesterday, we transformed ''E. coli'' DH5α with pSB1C3 and [http://partsregistry.org/Part:pSB1AK3 pSB1AK3] with [http://partsregistry.org/Part:BBa_B0014 BBa_B0014]. After checking the plates today for colonies we observed lots of pink colonies on the pSB1C3 plates and numerous white colonies on the [http://partsregistry.org/Part:pSB1AK3 pSB1AK3] with [http://partsregistry.org/Part:BBa_B0014 BBa_B0014] plates. Plates were then stored at 4°C as colonies will be used for minipreps.
 {{Team:Newcastle/footer}}

Team:Newcastle/27 July 2010

From 2010.igem.org