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=
-====Aim====
+[[Image:Newcastle alan chromosome.jpg|thumb|200px|right]]
-The aim of today's experiment is to extract genomic DNA from ''Bacillus subtilis'' strain 3610 and 168,genes from which will be needed for the swarming biobrick and ''rocF'' biobrick.
+[[Image:Newcastle ice chromosome.jpg|thumb|200px|right]]
+==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 [[Team:Newcastle/Swarming|swarming BioBrick]] and [[Team:Newcastle/Urease|''rocF'' BioBrick]] will then hopefully be obtained from the genomic DNA using PCR.
-====Procedure====
+==Protocol==
-* Please refer to [[Team:Newcastle/DNA extraction| DNA extraction of ''Bacillus subtilis'']]
+* Please refer to: [[Team:Newcastle/DNA extraction| DNA extraction of ''B. subtilis'']] for materials required and protocol.
-====Discussion====
+==Discussion==
-At the end of the DNA precipitation step, we did observe a small white pellet in all the eppendorf tubes.
+At the end of the DNA precipitation step, we observed a small white pellet in all the eppendorf tubes.
-====Conclusion====
+==Conclusion==
-Th experiment was successful and we would check the content and the purity of the extracted DNA by using PCR on 28th July, 2010.
+The experiment was a success! The quality of the extracted DNA will be checked by using PCR on 28th July, 2010.
-==='''Gel Electrophoresis'''===
+=Preparation for cloning of the rocF BioBrick=
-====Materials====
+==Results==
-[[Image:P7270470.JPG|thumb|right]]
-# 1% Agarose TAE
-# SafeView
-# TAE Buffer
-====Protocol====
+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.
-* Please refer to [[Team:Newcastle/Gel electrophoresis|Gel Electrophoresis]]
-====Discussion====
-* No band was observed on the gel.
-====Conclusion====
-The PCR did not work. It could be due to the DNA not being extracted or we 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.
 {{Team:Newcastle/footer}}

Team:Newcastle/27 July 2010

From 2010.igem.org