Materials and Methods
Restriction Enzyme Digestion – The experiment was begun after putting on gloves to avoid any chemical contact with the skin. Four microtest tubes were obtained, and each of them was labeled to contain the different enzymes or suspect DNA. Two of the microtest tubes were used for suspect one and the two different restriction enzymes, while two other microtest tubes were labeled for suspect two and the two restriction enzymes. After labeling the tubes, the contents that were at the bottom were taken out by slightly tapping them. Then to begin setting up the enzyme reactions, a micropipette was used to obtain 10 μL of the reaction buffer which was added to each of the four test tubes. The buffer is important because it carries the electrical current from the power supply in the gel. After the reaction buffer was in each, the microtest tubes were individually filled with their specific enzymes and DNA, shown in summary through Table 1.1 below. The restriction enzymes are used to cleave the DNA at specific
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The use of gel electrophoresis to separate DNA fragments by size and charge can be used in genetic testing, like paternity or maternity tests. This method could allow the DNA of the child to be accurately compared with the DNA fragments and banding patterns of one of the parents. This can allow one to determine the biological parents of a child. Another important use of DNA fingerprinting is the diagnosis of many inherited diseases. The DNA fingerprint of a person can allow doctors to determine if they carry an inherited disease through comparison of the band patterns, and if they are diagnosed, a treatment for the disease can be provided early on. DNA fingerprinting is extremely useful and essential in the improvement of lives through the medical field, as well as its other uses in solving crimes and cases of biological
It can be used in the real world to identify relationships between individuals by determining if two people are related. It could also be used to touch upon heritage from looking at back in generations. DNA fingerprints can also be used to identify a victim and help identify suspects in a crime scene. Lastly it can be used for personal identification. In some places, it requires you to give DNA fingerprints in order to identify a person.
1. From what organism are restriction enzymes derived? What role do these enzymes play in this natural host?Phages. Enzymes
Restriction enzymes cut DNA at certain sites to create multiple DNA fragments. Restriction enzyme HindIII has known DNA fragment lengths and recognition sites when digesting lambda DNA, while the lambda DNA recognition site for restriction enzyme XhoI is unknown. The goal of this study is to determine the lambda recognition site of XhoI by comparing a HindIII digest and a HindIII and XhoI double digest on an electrophoresis gel. The HindIII digest had a band at 9.4 kb, but this band was not visible in the double digest, therefore we concluded the recognition site for XhoI was around 9.4kb. There were also two additional DNA
After the substrate solution was added, five drops of the enzyme were quickly placed in tubes 3, 4 and 5. There were no drops of enzyme added in tubes 1 and 2 and in tube 6 ten drops were added. Once the enzyme solution has been added the tubes were then left to incubate for ten minutes and after five drops of DNSA solution were added to tubes 1 to 6. The tubes were then placed in a hot block at 80-90oC for five minutes. They were then taken out after the five minute period and using a 5 ml pipette, 5 ml of distilled water were added to the 6 tubes and mixed by inversion. Once everything was complete the 6 tubes were then taken to the Milton Roy Company Spectronic 21 and the absorbance of each tube was tested.
Analysis of DNA from practicals 1 and 2 using the technique of agarose gel electrophoresis and analysis of transfomed E. coli from practical 2 (part B)
DNA Fingerprinting, also known as DNA Profiling, is a method used to identify a person using DNA patterns that are specific to him/her. 99.9% of DNA is identical in every human being, but .01% is enough to distinguish between people. It is most commonly used in criminal cases to link a criminal to his/her crime scene, but is also used for paternity/maternity tests, and immigration records. Usually a skin, hair, or body fluid sample is collected from a crime scene or criminal or test candidate, then DNA is extracted and cut using enzymes that recognize patterns in DNA and run through a gel by an electric current in a process called electrophoresis (Annely).
The purpose of the experiment was to isolate plasmid DNA, followed by restriction digestion using restriction endonucleases and then visualizing the digested fragments after subjecting to gel electrophoresis. Plasmid DNA (pSP72 DNA) was isolated from Escherichia coli KAM32 (E.coli) cultures using the QIA prep miniprep kit and then subjected to restriction digestion by EcoRI and HindIII. The restriction digested DNA was then loaded into the wells of 0.7% agarose gel and subjected to electrophoresis. It can be concluded from our results that our plasmid DNA isolation was successful and the restriction digestion results were partially in agreement with our hypothesis.
Human DNA is very similar to one another, but only about 0.1% is different from the next person. That 0.1% can tell a person’s eye color, hair color, and other physical features. DNA analyst are able to take a drop of blood, the size of a dime, and duplicate the number DNA found in that drop. With the ability to duplicate DNA, analysts can have a back-up, in the event a human error were to occur. Analyst can tell you exactly where your ancestors came from and the percentage that is still inside your DNA. DNA is a very powerful tool that can identify a murder if the individual left any blood, saliva, skin tissue, hair or semen. The education needed to be able make use of the DNA consists of a great deal of science classes.
Deoxyribonucleic acid (DNA) has been used to analyze and prove innocence or guilt of suspects of crimes with great accuracy. DNA is part of everyday life. It is the heredity material in humans and almost all other organisms. While being part of an investigation. DNA has helped to solve crimes. There is a couple ways that DNA left behind can be tested to solve a crime. Either if the suspect has been caught and or had his or her DNA tested, or if he or she has left behind any biological evidence. Which then needs to be tested to see if it matches the DNA found in the crime scene to his or hers DNA. The result to this comparison may help establish if the suspect committed the crime.
DNA fingerprinting is a technique that is used to determine how likely it is whether genetic material came from a specific person or family group. Since 99% of human DNA is identical, that means that it is only 1% of our DNA which is different, and it is that 1% that we look at when we are attempting to determine the origin of a DNA sample.
The primary function of the digestive system is to transfer nutrients, water, and electrolytes from the food consume into the body’s internal environment. The ingested food is essential as an energy source, or fuel, from which the cells can generate ATP to carry out their particular energy-dependent activities such as contraction, transport, synthesis, secretion and even renewal of body tissues. Three primary categories of food ingested by humans which are carbohydrates, proteins and fats emerge as large molecules. These large molecules cannot cross plasma membranes intact to be absorbed from the lumen of the digestive tract into the blood or lymph; hence, it must undergo degradation in size (Sherwood, 2013). This
These strands give each any every person a unique genetic code, different from the rest of the world. “99.9% of the DNA from two people will be identical. The 0.1% of DNA code sequences that vary from person to person are what make us unique,” (BBC. 2013). The advancements started in the late 1900’s. Scientists were using programs to help identify DNA. Eventually a forensic board opened in 1990, setting a standard of laws and standards that the new base of technology would have to follow. DNA identifying is a cellular biology process. It takes place by using a sample of DNA, usually one from the potential criminal found at the crime scene or a victim's body, then “the DNA is isolated from the cells and millions of copies are made, using a method called 'polymerase chain reaction', or PCR. PCR uses a naturally occurring enzyme to copy a specific stretch of DNA over and over again. Having lots of DNA makes the genetic code easier to analyse. The DNA molecules are then split at particular locations to separate them into known 'chunks' and the code at those specific points is analysed to create a DNA fingerprint,” (BBC, UK 2014). DNA forensic technologies have made some very important advancements. Such as freeing David Vasquez, a wrongly convicted man of one of serial killer Timothy Wilson Spencer’s crimes. DNA testing has also made historical advancements proving that Anna Anderson who had famously claimed to be the Grand Duchess Anastasia, the only survivor of the Russian royal family. After Anderson died, DNA tests proved she was no relation to the Imperial family.Forensic testing is usually broken down into two ways. First is Y-marker analysis, used when men are involved and often sexual assault cases. As its name suggests, this technique examines several genetic markers found on the Y chromosome. Scientists look into the Y gene and
Plasmid DNA with Restriction Digest: The purpose of restriction digest of plasmid DNA is to understand how each DNA plasmids was cut with the given restriction enzymes and perform gel electrophoresis to observe the samples. Nine restriction digests were created, containing three digests for each of the three plasmid DNAs identifying as recombinant, non-recombinant, and unknown. Out of the nine digests, six are actual digests and three are undigested controls. A master mix is created to add to each of the nine samples with its following stock ingredients: 10 ul of 2X Reaction Buffer, 1 ul of Nco1, X ul of sterile water (Single digest), 10 ul of 2X Reaction Buffer, 10 ul plasmid DNA, 1 ul Nco1, 1 ul of Not1, and X ul of sterile water (Double
The process of DNA fingerprinting in humans involves the replication and arrangement of extracted DNA, to create a pattern/fingerprint that is viable for comparison. This process involves the application of DNA extraction, digestion by restriction enzymes, Polymerase Chain Reaction (PCR) and gel electrophoresis. This results in a DNA profile with bands of varying widths that can be used for the comparison of genetic information. DNA extraction occurs in three stages. Firstly, a