Latest revision as of 19:16, 25 October 2010

Research

Today we initiated with our research in urease which would lead to calcium carbonate production inside the crack.

Wray LV, Ferson E and Fisher SH. (1997). "Expression of the Bacillus subtilis ureABC operon is controlled by multiple regulatory factors including CodY, GlnR, TnrA, and Spo0H". Journal of bacteriology. 179(17). 5494-501.
Kim JK, Mulrooney SB and Hausinger RP. (2005). "Biosynthesis of Active Bacillus subtilis Urease in the Absence of Known Urease Accessory Proteins". Journal of Bacteriology. 187(20). 7150-7154.

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-===Transformation===
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-# 1:3
+=Research=
-# 1:5
-# Vector (no insert), negative control for ligation
-# DH5Alpha (without plasmid)
-# DH5Alpha (with plasmid, pSB1AT3)
-Protocol:
+Today we initiated with our research in urease which would lead to calcium carbonate production inside the crack.
-# Thaw a 200µl aliquot of E. coli DH5Alpha. Add the transforming DNA. We added 5µl of vectors from tubes 1 to 3 and 1µl of vector from tube 5 because vector  from tube 5 is has not been ligated whereas tubes 1 to 3 have been ligated and will contain cells in different forms.
+#Wray LV, Ferson E and Fisher SH. (1997). "Expression of the Bacillus subtilis ureABC operon is controlled by multiple regulatory factors including CodY, GlnR, TnrA, and Spo0H". ''Journal of bacteriology''. 179(17). 5494-501.
-# Incubate for 45 min on ice.
+#Kim JK, Mulrooney SB and Hausinger RP. (2005). "Biosynthesis of Active Bacillus subtilis Urease in the Absence of Known Urease Accessory Proteins". ''Journal of Bacteriology''. 187(20). 7150-7154.
-# Heat-shock the cells at 42°C for 120 secs, and place on ice again for 3-4 min.
-# Add 1ml of LB broth to each tube and incubate the cells at 37°C for 1.5 hr (static for initial 20 mins, shaking for remaining time).
-# Plate out 200 µl/plate on LB (agar at 1.5%), with ampicilin.
-# Incubate plates overnight at 37°C.
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