a genomic library‚ genomic DNA from Vibrio fischeri was first isolated then treated with Sal I restriction enzyme to generate inserts (smaller fragments of DNA). Sal I restriction enzyme was also used to treat the vector plasmid in order to digest the V. fischeri DNA fragments. The inserts and the vector were then ligated together. E.Coli cells were then made competent in order to take up the plasmid DNA by transforming these competent cells with a “ligation mixture to create a population of host
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|Regional & State |Regional & State |National (all topics) | |DNA structure & function |Lac & Trp Operons |Restriction mapping | |DNA Semi-conservative Replication |DNA Fingerprinting/RFLP |Mitochondrial DNA | |Gene expression (transcription and translation
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Chapter 9 DNA-Based Information Technologies Multiple Choice Questions 1. DNA cloning: the basics Page: 307 Difficulty: 1 Ans: C Restriction enzymes: A) act at the membrane to restrict the passage of certain molecules into the cell. B) are highly specialized ribonucleases that degrade mRNA soon after its synthesis. C) are sequence-specific DNA endonucleases. D) are very specific proteases that cleave peptides at only certain sequences. E) catalyze the
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Isolated DNA Products Amplified Via Polymerase Chain Reaction and Cloned Biotechnology: DNA WPUNJ December‚ 2012 Abstract Isolated DNA from mouse‚ plants‚ and plasmid DNA were used for Polymerase Chain Reaction (PCR) for DNA amplification. The purpose of this experiment was to study the success rate or optimization of PCR of DNA‚ using both manual and kit methods. This set of experiments gives an insight to the relative difficulties associated with the optimization of a variety
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Cloning of plasmid pUC19 in E.coli bacteria Introduction One aspect of the DNA cloning experiments that is carefully considered is the selection of cloning vectors. A variety of vectors have been created‚ each being suitable for a particular use. One common vector used in laboratories is a plasmid called pUC19. It is 2686 base pairs long and possesses an origin of replication which allows the production of over 100 copies in a competent E.coli cell. It possesses a multiple cloning site (MCS)
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forms a double helix. Restriction enzymes are bacterial enzymes that cut both strands of DNA at specific nucleotide sequences. These enzymes digest DNA by cutting the sequence at specific locations called restriction sites. Some restriction enzymes cleave DNA strands in the center of the restriction site‚ while others cut the backbone in two places‚ so the pieces have single-stranded sticky ends of unpaired nucleotides. The two pieces of DNA that are cut with the same restriction enzyme can be “pasted”
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Are You a Good or Bad Student? Being a good student is what everyone wants to believe they are. But in reality we all know that there are bad students. I‚ myself would like to believe I am a good student but when I looked over the facts it seems that I am not a bad or a good student. The first and foremost important quality of a good student is‚ of course‚ hard working. We can’t have a good result in academic success without training and effort. The next quality is active in community. A good student
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determining if any genetic transformation has occurred. By combining +pGLO LB and ampicillin we should get an ampicillin resistant gene and by using –pGLO we should create a non-genetic resistant bacteria. The pGLO plasmid has the GFP (green fluorescent protein) gene and the gene that allows the plasmid to be resistant to the antibiotic ampicillin. The most important part of this experiment is the “heat shock treatment” because the E. coli membrane becomes permeable and increases the competency of the cells
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b. Electroporation – shock DNA to open pores in plasma membrane so DNA can enter cell c. Have small DNA molecules w/ 0 plasmids 1 ori and carry accessory genes 3) Restriction endonucleases attack DNA molecule from inside. Exonucleases – chop off 1 nucleotide at a time at the ends of the DNA molecule. Not in abundant in prokaryotes Bacterial DNA gets methylated – restriction enzymes cut only unmethylated DNA Bacterial DNA gets bacteria add CH3 to Cytosine or Thymine to protect their own enzymes
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Restriction Fragment Length Polymorphism (RFLP) Restriction Fragment Length Polymorphism (RFLP) Definition Definition In molecular biology‚ restriction fragment length polymorphism‚ or RFLP is a technique that exploits variations in homologous DNA sequences. It refers to a difference between samples of homologous DNA molecules that come from differing locations of restriction enzyme sites‚ and to a related laboratory technique by which these segments can be illustrated. In RFLP analysis
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