Dna Isolation Lab Report

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.

DNA samples are submitted to a certified laboratory and undergo the following process (DNA Evidence. n.d.): • Extraction is the process of releasing the DNA from the cell. • Quantitation is the process of determining how much DNA you have. • Amplification is the process of producing multiple copies of the DNA in order to characterize it. • Separation is the process of separating amplified DNA product to permit subsequent identification.

In 1948, Linus Pauling discovered that many proteins take the shape of a helix. At Cambridge University, James Watson and his research partner Francis Crick had become interested in Linus Paling’s work. Their approach was to try to make a physical model of what DNA looks like to narrow down the possibilities and eventually create an accurate model of the molecule. In 1951, Watson attended Rosalind Franklin’s lecture on the current work that she had done.

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.

There are three kinds of DNA with specific functions. First, messenger RNA (mRNA) is responsible for transcribing the information from the DNA. That information is then sent to the ribosomes and cytoplasm. The second, transfer RNA (tRNA) transfers amino acids to the mRNA in a ribosome.

DNA Profiling/Database: The concept is such that, DNA samples are taken from an individual and it is analysed in a laboratory. Further, based on this analysis,

In Fred Sanger Method, the DNA to be sequenced serves as a template for DNA synthesis. A DNA primer is needed which is designed to be a starting point or the initiating point for DNA synthesis on the strand of the DNA to be sequenced. The primer is hydrogen bonded to the 3 ' end of the DNA to be sequenced. The DNA with the primer is divided into four separate reaction mixtures. Each reaction mixture contains all four dNTPs and in addition, one of the four dideoxy analogs (dideoxyribonucleoside triphosphates ddNTPs) of the deoxyribonucleoside triphosphates.

Deoxyribonucleic acid (DNA) is a molecule found in all forms of life that is passed down from parents to offspring. What makes each DNA unique is the chemical makeup of the molecule sometimes referred to as the “blueprint of life.” (BIO). DNA is made up of nucleotides consisting of a sugar, a phosphate and a base pair. About six million nucleotide base pairs make up DNA in each cell.

After his discovery other method were found and that’s why RFLP is used rarely. How is DNA applied? There are three methods of DNA profiling: RFLP, PCR and STR. RFLP: DNA is cut by restriction enzymes according to repeated sequences of DNA bases, which is unique for every person.

Polymerase Chain Reaction (PCR) is a laboratory technique used in molecular biology to generate a small section of DNA or genes, PCR uses primers to amplify specific genomic DNA sequences with the help of special enzymes, PCR process uses short sequences of DNA and primers and selects specific chromosomes of DNA for replication. (McPherson and Møller, 2009) In addition, there are three main steps involve PCR process, first step of PCR is denaturing when DNA is heated to 90-95 degrees Celsius and double DNA strand is separated in to two single strands DNA, these single strand DNA will act as a template for the new strand DNA, because of the higher temperature the hydrogen bond which holds the complementary strands of DNA together is broken down at this step. (Wheeler et al., 2004) The second stage in the PCR cycle is annealing, this step the temperature is cooled to 50-56 degrees Celsius so the DNA primers can attach to the template DNA by the hydrogen bonding.

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.

The same region is also amplified on both chromosomes, however they are different sizes, which are then put into gel

Furthermore, Acinetobactor baylyi ADP1 like most organisms undergoes a process known as DNA recombination, where two complementary DNA strands cross and exchange portions of DNA. During recombination, a structure known as a Holliday Junction forms and must be resolved, completing the exchange of DNA (Aravind et al. 2000). Recombination is a crucial mechanism in both gene amplification and deletion. Specifically, ADP1 contains a protein called YqgF, a putative Holliday Junction Resolvase, due to its structural similarity to a known resolvase named RuvC (Aravind et al.

My experience for the stakeholder’s meeting and writing my paper was different. For example in the stakeholder’s meeting we discussed the situation if golden rice should be given to children in public schools based on our roles, which was different to the paper because it was more about the facts and your role wasn’t a part of it. For the paper I thought having to find out how golden rice was created was challenging, and for the stakeholder’s meeting I thought talking about if children should eat golden rice in public schools was challenging. The benefit for each is that I learned new information. Studying golden rice really impacted me when we learned about DNA.