3 research outputs found
Optimizing Genetic Manipulation of Methanogens through Faster Cloning Techniques
No full textMethanogenesis is the biological production of methane. Only anaerobic archaea known as methanogens are capable of such a metabolic feat. They have strict living conditions and substrate sources which determine their rate of metabolism. This is of particular importance from a greenhouse gas reduction perspective or biogas capturing perspective. One of the best ways to optimize methanogen methane production is via genetic manipulation. The current procedures are timely though, therefore a faster cloning processes should be developed. The objective of this study was to optimize a premade genetic transformation kit known as the Gibson Kit. The Gibson Kit was supposed to ease the work of genetic manipulation by combining several linear chunks of DNA together at once via the use of sequence overhangs. The resulting plasmid from the Gibson could then be transformed into E. coli where E. coli is supposed to replicate the plasmids extremely quickly. Afterwards, that plasmid could then transform methanogens. Several baseline PCR (polymerase chain reaction) transformations were performed to linearize and amplify the desired plasmid of pNB72.3. PCR amplifications of the desired gene segments which would be added to the plasmid were also performed. The desired gene segments being assembled were supposed to take out the production of Cytochrome C within the methanogen electron transport system by deleting the ccmf gene. Several PCR experiments were carried through without success. The cause of the failures included the primers “hair pinning” at the recommended annealing temperatures, primers being too nonspecific, and primers unable to perform efficiently at the recommend annealing temperatures. After several tries of no success, the Gibson Kit was tested without PCR linearization of the circular plasmid. Instead, standard digestion was relied on. With standard electro competent cell transformation and antibiotic screening, the Gibson product was then tested for successful transmission of the plasmid to the E. coli. Results were negative; therefore, the optimization of faster cloning techniques was unsuccessful, but it will help guide future efforts
PENGARUH KECERDASAN EMOSIONAL DAN PENGEMBANGAN SUMBER DAYA MANUSIA TERHADAP KINERJA KARYAWAN PT CENTRAL BATAM RAYA
No full textEmployee performance has a very important role because it will affect the employee output so that will affect the performance of a company. This study analyzes employee performance at PT Central Batam Raya are reviewed through the variables of emotional intelligence and human resource development. This research is a quantitative research with the method used is descriptive analysis and multiple regression analysis and data collection techniques through questionnaires. Questionnaires distributed amounted to 102 respondents. The method used in this study was chosen mainly because of the limited time, cost and energy that the author has, so that in this case the writer hopes to be able to examine specific aspects of a social situation in depth. Data collection is done by primary data and secondary data. The results showed that the variable emotional intelligence and human resource development had a significant effect simultaneously on employee performance with t count> t table (4,860 > 1,98397) and (4,935 > 1,98397), significant value <0.05 and F count> F table (70,337 > 3,09) with a significant value <0.05
