Latest revision as of 19:16, 25 October 2010

Home

Team

Problem

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.

@@ Line 1: / Line 1: @@
-===Transformation===
+{{Team:Newcastle/mainbanner}}
-Link to transformation protocol (to be added)
+=Research=
+Today we initiated with our research in urease which would lead to calcium carbonate production inside the crack.
-====Tubes used:====
+#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.
-Each of the following five tubes contain 200 µl of competent ''E. coli'' DH5alpha. To this the DNA to be transformed was added.
+#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.
-# 1:3 (contains LacI insert and pSB1AT3 vector in the proportion of 1:3)
-# 1:5 (contains LacI insert and pSB1AT3 vector in the proportion of 1:5)
-# Negative control for ligation (contains vector with no insert)
-# Control for transformation (without plasmid)
-# Control for transformation (with plasmid, pSB1AT3)
-====Protocol:====
-# Thaw a 200 µl aliquot of ''E. coli'' DH5alpha and add the transforming DNA. For tubes 1-3 5 µl of vector was added, for tube 4 no vector was added and for tube 5 only 1 µl of vector was added. This is because tubes 1 to 3 have been ligated and will contain cells in different forms of the ligated vector whereas tube 5 contains the ligated vector (our control).
+{{Team:Newcastle/footer}}
-# Incubate for 45 mins on ice.
-# Heat-shock the cells at 42°C for 120 secs, and place on ice again for 3-4 min.
-# Add 1 ml 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.

Team:Newcastle/9 July 2010

From 2010.igem.org

Latest revision as of 19:16, 25 October 2010

Research