This experiment tested the growth of E.coli with inserted plasmid on an agar plate with Ampicillin. One colony of E.coli resistant to Ampicillin was grown during this experiment. The overall goal of the experiment was to successfully grow E.coli on the agar plate‚ which would show that the plasmid had been effectively inserted into the bacteria’s genes. This experiment helped students understand how plasmids were inserted into bacteria and used in real life situations. It also showed how the bacteria’s
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The pGLO plasmid‚ which has the bla and GFP gene‚ was developed so it can operate like the arabinose operon and be able to transcribe genes in the presence of arabinose sugar. In this plasmid‚ the cluster of genes is regulated by the spontaneous on/off element by a single promoter‚ which is dependent on the DNA binding protein araC. This protein is at the binding site for RNA polymerase at the beginning of the operon‚ so when arabinose is present (like in one of the experimental plates)‚ it is taken
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NA transformation of E. coli: The two plasmids were added to individual tubes containing E. coli and one with no plasmids. The three samples of E. coli were heated in a 42°C water bath for 90 seconds to heat shock the bacteria so that the plasmids would be taken up by the E. coli. These samples were then incubated at 30°C for half an hour and then plated on LB agar. Each tube was plated on an LB plate and a LB + ampicillin plate. Ampicillin is an antibiotic that is effective against E. coli‚ both
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Molecular Biology Lab Report Payton Jackson Introduction In this lab‚ I am going to use antibiotic-resistance plasmids to transform Escherichia coli. Materials For this lab you will need the following: LB Agar Petri dishes Beakers Test tubes CaCl2 solution Sensitive E. coli (-ampR) amp plasmids ampicillin -amp cells Water bath to heat shock cells A freezer to incubate cells Process Step 1: Wash hands and sanitize lab setting. This will prevent anything reacting with a
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using calcium chloride/heat shock. This is done by incorporating the plasmids into chemically competent cells that were made permeable by the calcium chloride solution and heat shock. In 1928‚ Frederick Griffith‚ a physician from London‚ was he first person to experiment with bacterial transformation. He permanently transformed a safe‚ nonpathogenic bacterial strain of pneumococcus into a deadly pathogenic strain. [1] Plasmids are small circular autonomously replicating pieces of DNA that can be
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uses of plasmids in G.M. experimentation. Plasmids are extrachromosomal genetic elements found in a variety of bacterial species. They are double stranded; autonomously replicating‚ supercoiled‚ covalently closed circular (ccc) DNA molecules that range in size from 1 kb to greater than 200 kb. Often‚ plasmids contain genes coding for enzymes that‚ under certain circumstances‚ are advantageous to the bacterial host (Table 1). Table 1. Some of the phenotypes conferred by different plasmids that
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Rainbow Transformation1 lab manual. An Escherichia coli bacterial reference plate was used to obtain colonies which were resuspended into a CaCl2 solution that was previously kept on an ice bath. The rainbow transformation mixture containing the plasmid DNA was then added to half of the E. coli cells. These cells were later placed into a water bath set to 42ºC and “heat shocked” to promote the entrance of DNA into the cells. Moreover‚ a Recovery Broth was added to the sample and the sample was left
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process which involves plasmid DNA being bound to the cell surface and the subsequent uptake of DNA by the cell (Panja et al.‚ 2008). For artificial transformation of E. coli cells with plasmids‚ plasmid DNA has to be extracted from bacterial cells using the High-Speed Plasmid Mini Kit‚ which is then mixed with competent E. coli cells followed by heat shock and the streaking of transformed cells on two different types of agar plate (LB and LB+ampicillin). The extracted plasmid DNA is important as it
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Detection of plasmids in arsenic (As) resistant bacteria isolated from As-contaminated groundwater Mahima Rani and Pinaki Sar* Department of Biotechnology‚ Indian Institute of Technology‚ Kharagpur‚ West Bengal–721302‚ India mahima06bt11@gmail.com‚ Ph: +91 8348523016 Abstract Role of plasmids in conferring resistance to several heavy metals and antibiotics to naturally occurring bacteria is well known. In contaminated environments‚ presence of metal resistant genes on plasmids often provide
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of this experiment‚ we transformed the bacteria into an antibiotic resistant form by inserting a plasmid into it. We used heat shock in order to make the bacteria capable to uptake a plasmid in the presence of calcium ions that help disrupt the cell membrane (heat shock is the combination of altering hot and cold). When they are capable of accepting plasmids‚ the bacteria are incubated with plasmids that carry the resistance to a particular antibiotic‚ in this case ampicilin. We also ran a control
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