Latest revision as of 15:08, 27 October 2010

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Problem

Genomic DNA extraction from Bacillus subtilis strain ATCC 6633

Aim

The aim of this experiment was to extract genomic DNA from Bacillus subtilis strain ATCC 6633. This strain has the subtilin genes, required for our Subtilin Production and Immunity parts.

Protocol

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

Results

Results were as expected see: https://2010.igem.org/Team:Newcastle/7_July_2010

Conclusion

See: https://2010.igem.org/Team:Newcastle/7_July_2010

Research

Today we also continued with our research on filamentous cells which will act as reinforcing fibres in the crack.

Baarle S van and Bramkamp M. (2010). "The MinCDJ system in Bacillus subtilis prevents minicell formation by promoting divisome disassembly". PloS one. 5(3). e9850.
Mo AH and Burkholder WF.(2010). "YneA, an SOS-induced inhibitor of cell division in Bacillus subtilis, is regulated posttranslationally and requires the transmembrane region for activity". Journal of bacteriology. 192(12). 3159-73.

@@ Line 1: / Line 1: @@
-'''[[Gram-positive Bacteria Chromosomal DNA Extraction Protocol]]'''
+{{Team:Newcastle/mainbanner}}
-===Puregene DNA isolation===
-# Grow overnight 7.5ml cultures in THYB containing 30&#0956;g/ml hyaluronidase.
-===Cell Lysis===
+=Genomic DNA extraction from ''Bacillus subtilis'' strain ATCC 6633=
-# Two sets of chromosomal DNA were being done.
-# Cells go through 3600 x g centrifugation for 15 minutes to pellet.
-# The supernatant in the tubes are poured away.
-# 0.5ml of cell suspension solution is added to the eppendorf tubes. The solution in the tubes are pipetted up and down to resuspend cells. The solution is then transferred to another 1.5ml tube.
-# 50&#0956;l of lysozyme and 7.5&#0956;l of lytic enzyme solution is added into each tube. The tubes are then inverted 25 times to mix the solution.
-# We then incubate them at 37&#0176;C for 30 minutes in order to digest the cell walls. The tubes are inverted occasionally during incubation.
-# To pellet the cells, we centrifuged it for 10 minutes and poured away the supernatant after that.
-# 0.5ml of cell lysis solution is added to the pelleted cells and the solution is pipetted up and down in order to lyse the cells. The samples are then heated for about 5 minutes at 80&#0176;C.
-===RNase Treatment===
+==Aim==
-# 6&#0956;l of RNase A solution is added into the cell lysate.
+The aim of this experiment was to extract genomic DNA from ''Bacillus subtilis'' strain ATCC 6633. This strain has the subtilin genes, required for our Subtilin Production and Immunity parts.
-# The tubes are inverted 25 times in order to allow the solutions to mix. It is then incubated at 60minutes at 37&#0176;C.
-===Protein Precipitation===
-# Samples are cooled on ice.
-#
+==Protocol==
+Please refer to: [[Team:Newcastle/DNA extraction|DNA extraction of ''Bacillus subtilis'']] for materials required and protocol.
+==Results==
+Results were as expected
+see: https://2010.igem.org/Team:Newcastle/7_July_2010
+===Conclusion===
+See: https://2010.igem.org/Team:Newcastle/7_July_2010
+=Research=
+Today we also continued with our research on filamentous cells which will act as reinforcing fibres in the crack.
+#Baarle S van and Bramkamp M. (2010). "The MinCDJ system in Bacillus subtilis prevents minicell formation by promoting divisome disassembly". ''PloS one''. 5(3). e9850.
+#Mo AH and Burkholder WF.(2010). "YneA, an SOS-induced inhibitor of cell division in Bacillus subtilis, is regulated posttranslationally and requires the transmembrane region for activity". ''Journal of bacteriology''. 192(12). 3159-73.
+{{Team:Newcastle/footer}}

Team:Newcastle/6 July 2010

From 2010.igem.org