DNA after synthesis were PCR amplified using pFx enzyme...

DNA after synthesis were PCR amplified using pFx enzyme respectively for both premature AI and mature AI. The products of the PCR have blunt ends due to the amplification done by pFx. Mature AI was not able to be amplified using pFx so it was done using Taq Polymerase. PCR clean up of the products were done and was quantified using Nanodrop. Restriction digestion of the Mature AI, Premature AI and pET28-a was done at the Xho and Not1 restriction sites. Purification of the digested products were done using Sodium acetate precipitation protocol. Ligation of the purified products were done using Ligase enzyme. Stage 1 Stage 2 (Cycle 33) Stage 3 95⠁ ° C 95⠁ ° C 55⠁ °C 68⠁ °C 68⠁ °C 4⠁ °C 2:0 min 30Sec 45Sec 1:30min 7:0 min ∞ Table.†¦show more content†¦Pellet was resuspended in buffer and then SDS-PAGE was run for the samples 3.5.5. ProBond purification of Premature AI and Mature AI: Lysis and Suspension buffer composition for BL21 E.Coli cells protein expression are: †¢ Lysis buffer(pH 7.0): 50mM MOPSO, 10% Glycerol, 10mM MgCl2 †¢ Wash buffer(pH 7.0): Add 20mM Imidazole to the lysis buffer. †¢ Elution buffer(pH 7.0): Add 250 mM Imidazole to the lysis buffer. To remove precipitate containing salts column was run (affinity chromatography).In affinity chromatography 1 ml raisin was taken which contained nickel ions. It was equilibrated against tris buffer of pH 8. The equilibrated raisin was added in protein suspension then it was kept for binding for 2 hours. After the binding process, the column was run i.e. affinity chromatography was performed. Histidine tags were present in the protein, it has affinity for nickel ion, and so all Histags got binded with nickel. The elution fractions were collected which contained purified protein and its activity was checked and SDS-Page was run. Before adding resin sample was taken out for assay. After bound sample was taken out in separate tube for assay. About 37 mL of the wash buffer was used to wash the undesired proteins. First two and the last two washes were saved for the activity. About 4 mL of the elution buffer was used for eluting the desired protein. All fractions were

Ethical Issues Of Social Media - 963 Words

Ethical Issues in Social Media Emerging Information Technologies are faced with numerous ethical issues. One of the most centers of ethical debate in Information Technology is social networks. Social networking has changed from a niche phenomenon to mass adoption. Although the concept of social networking was formulated in the 1960s, the advent of fast internet has sent the concept viral. Currently, social networks have been dogged with myriad ethical issues. Privacy issues are the epicenter of these ethical issues. Some of the ethical problems that have emanated from social networks include hacking, stalking, credit card theft, and social network identity theft. Resultantly, most of social media users are haunted by the dilemmas such as how much personal information to put on social media, how they can control the audience of their profiles, and whether the information they delete from their profiles is well disposed of. 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Electric vs Light Microscopes free essay sample

What are some advantages and disadvantages of a light microscope and electron microscope? Some of the disadvantages of the light microscope include, that since it send light, the light waves are diffracted as they pass through the matter. Because of this, light microscopes can only produce clear images of objects to a magnification of about 1000 times. Another problem is that since most living cells are nearly transparent, you can’t see them. So you have to use dyes to stain them. The advantage of using a light microscope however is that you can use it to see cells and cell structures as small as 1 millionth of a meter! Electron microscopes are used to study things even smaller such as viruses or DNA molecules. Some disadvantages include the fact that while using a transmission electron microscope, its beams of electrons can only pass thorough thin samples, cells and tissue must be cut into ultrathin slices before being examined because of this. We will write a custom essay sample on Electric vs Light Microscopes or any similar topic specifically for you Do Not WasteYour Time HIRE WRITER Only 13.90 / page Images often appear flat and two dimensional also. Another disadvantage is that, electrons, unlike light, don’t come in colors. Scientists use computer techniques to add â€Å"false colors† because of this to make certain structures stand out. Louis Pasteur Louis Pasteur refuted the notion of spontaneous generation. He conducted an experiment by making to flasks that had â€Å"s† shaped tops. He put broth into the flasks, and then he heated them (boiled) to kill all existing bacteria. People assumed that if you left the flasks out that bacteria would grow or be â€Å"spontaneously generated† in the broth. It did not however, because the bacteria in the air could not go up the â€Å"s† on the flask because of gravity. He broke some of the â€Å"s† shapes of the ends of the flask and left some covered and then waited a few days. After a few days had passed, he checked the broths and only that of the broken flask had bacteria in it.