Thermo Scientific
DreamTaq™ Hot Start Green PCR Master Mix by Thermo Scientific
K9021
Info: Supplier Page
I conducted research on a peptidoglycan (PG) hydrolytic enzyme and demonstrated its unusual substrate specificity. This enzyme contributes to the PG sacculus expansion, highlighting the crucial role of cross-link cleavage in bacterial PG synthesis. Additionally, I explored the physiological function of orthologs from two model organisms, Caulobacter crescentus and Klebsiella pneumoniae. To achieve this, I cloned the orthologs from these bacteria under an IPTG-inducible promoter in the pTRC99a physiological vector. For the identification of positive clones, I extensively utilized the DreamTaq™ Hot Start Green PCR Master Mix to screen numerous colonies.
Screening colonies for positive clones.
E. coli cells
For colony PCR, dissolve a colony in 20 µl of water (making it viscous), and use 1 µl as a template for a successful PCR reaction.
Using this kit, I identified numerous positive clones encoding cell wall hydrolase orthologs, which I subsequently utilized to assess the extent of complementation in the E. coli cell wall hydrolase mutant. I demonstrated that orthologs from various bacteria effectively complement the growth defects observed in the E. coli mutant lacking its cell wall hydrolase.
30940749
Simple yet robust!
None.
Great product. I would definitely purchase this again. The perfect companion for bacterial geneticists.