Lab 3 – DNA extraction and visualization Journal -Madhu Thalari. 1.Describe the laboratory exercise as you interpreted it.? Ans: This lab has given me methods to extrct DNA from both plant cells and animal cells. The main steps that are followed in both methods made me understand the reasons behind them. In order to extract DNA we need break the barriers(cell wall and cellmembrane), remove water, protiens and other unwanted material, make sure that the chemical we used should not damage DNA that we need and add flouroscent material to visualize the DNA.
For this reason the DNA is denatured into single strands by incubation with sodium hydroxide solution (NaOH). If the DNA fragments are larger than 15kb, the gel can be treated with acid such as dilute HCl prior to blotting. The DNA is depurinated by the acid and breaks down into smaller pieces which allows more efficient transfer from the gel to the membrane. The Southern Blot Method begins with the DNA fragments being transferred to a nitrocellulose membrane.
Western blotting is a procedure used to detect specific proteins in a given sample. Gel electrophoresis is used to separate the proteins which can be observed as thick and thin bands on the electrophoresis gel. In this experiment we use SDS-free polyacrylamide gel. Sample proteins used in this case are bovine serum, human serum, goat serum, chicken serum and horse serum. Since the SDS is negatively charged, sample proteins move to the positively charged anode through the gel.
An example is sulfomethoxazole [SMX] of the sulfonamide family: some bacteria utilize para-amino benzoic acid[PABA] a start-up product in producing folic acid –containing intermediates for DNA replication, using the enzyme dihydroptorate synthase to produce dihydroptorate. SMX blocks this enzyme, but these days, study has shown some bacteria that totally for-go this PABA pathway, these bacteria are now resistant to SMX because it really has nothing to work on. Enzymatic destruction of antibiotics: some microbes develop antibiotics resistance by producing enzyme to destroy the antibiotics. An example is the beta-lactam antibiotics, namely penicillins, amoxicillin. These antibiotics have this part of their chemistry, the beta-lactam rings, some organisms especially the gram-negatives carry in their periplasm enzymes called beta-lactamses, to destroy any drug with this beta-lactam rings. Reduced uptake: some microbes also develop a mechanism to reduce the uptake of antibiotics, an example is resistance to the
Anti-infection agents are utilized to treat diseases caused by bacteria mostly Gram-negative bacteria. Nalidixic acid works by killing the microbes that are causing a contamination or infection. It does this by entering the bacterial cells and restraining a bacterial protein called DNA-gyrase. This catalyst is associated with duplicating and repairing the genetic material (DNA) of the microbes. On the off chance that this enzyme doesn't work, the microbes can't replicate or repair themselves and this executes them.
Water and Sewage Microbiology: 1. List the steps of in a water purification plant. a. Screening to separate the large contaminants from the water b. Coagulation to attract small contaminants c. Sedimentation where water sits and finishes coagulation d. Filtration to remove any small remaining contaminants and particles e. Disinfection by disinfecting chemicals such as chlorine to kill microorganism or remaining bacteria 2.
CCCCC We can use the following analogy to (Peter Daempfle, 2001) translate the DNA to mRNA. We can also use this analogy to determine from the mentioned sequences which is not a correct translation of the mRNA.DNA matches with mRNA in:A matches with UT matches with AG matches with CC matches with G The odd one out from the sequence is clearly (b) that is UTTCTTT this is because the code T is from a DNA sequence rather than from a mRNA sequence. Peter Daempfle (2001). `Essential Biology An applied approach` Kendall hunt publishing Company Correct Answer: n/a ********************************************************************************************************** 9. Some antibiotics are used to kill bacteria by stopping the ribosome from functioning.
This is another way to destroy the bacteria. For example, the antibiotics that name are Lenkomaisen and Macrolede. Another type of antibiotics is which attack the cell membrane. Such as in Benslimit and Bastrsin. The antibiotics are one of the most useful thing that the scientists discovered.
Chlorine dioxide responds specifically with amino acids and the RNA in the cell. It is not clear whether chlorine dioxide attacks the cell structure or the acids inside the cell. The generation of proteins is avoided. Chlorine dioxide influences the cell layer by changing film proteins and fats and by anticipation of
The purpose of this experiment is to create a complete genomic library of Aliivibrio fisheri through the use of the lux operon. The examination of the lux operon gene occurs through the extraction of the DNA of Aliivibrio fischeri and digest a large piece of DNA to smaller random pieces. The fragment of DNA will later be ligated together in plasmid. Plasmid acts as vectors to transport DNA from one organism to another. The DNA will then run through a UV-visible spectrophotometer to test the absorbance of the extracted DNA.
Transformation in bacteria usually takes place when a bacterial cell accepts strange DNA and integrates to its own DNA. The transformation normally takes place within plasmids, which are tiny circular DNA molecules that have been separate from its own chromosome. The copies of the same plasmid range from 10 to 200 copies within a cell. These copies of plasmids may multiply when the chromosome replicate or multiply independently. One plasmid has a range of 1,000 to 200,000 base pairs.
n this lab, there were four objectives needed to be met. The first one was to perform a genetic transformation procedure, the second was to move genes from one organism to another using a plasmid as a vector, and the third was to manipulate tools of biotechnology. The bacteria E. coli was used to manipulate and transform. The E. coli would be tested for ampicillin resistance and a green fluorescent glow. One hypothesis made for this lab was that the bacteria that developed a resistance to ampicillin would reproduce even in the presence of the ampicillin.