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== | ==Electrophoresis Day== | ||
A Flower of A Different Color | A Flower of A Different Color | ||
Today, like any other lab day, Angie, Diwash, Sushma and Binu, again, were working on the plasmid extraction hoping to get good results. Angie and Diwash worked on the plasmids that were extracted on 7th Sep., ie. restriction digestion with EcoR1 and the electrophoresis of the same. Sushma and Binu did the plasmid extraction with the cultures that was prepared yesterday (from MIT), for backup. The results of the gel electrophoresis was disappointing, again, as we didnt get the bands of the right size. But this gel also showed that the problem couldn't lie with the plasmids alone, as even pBluescript showed some bands of the size that was not expected. We along with Axel came to the conclusion that it probably was not just the plasmids but also the restriction enzyme, EcoR1 that was messing up the experiment. There was a slight possiblity that even RNAse could be the culprit. So,it was decided that on Tuesday, we will be digesting the plasmids (the third set and the one that was extracted today)with 3 restriction enzymes - new EcoR1, and the enzymes that would help to insert the gene into the plasmid, ie. Spe1 and Xba1. | Today, like any other lab day, Angie, Diwash, Sushma and Binu, again, were working on the plasmid extraction hoping to get good results. Angie and Diwash worked on the plasmids that were extracted on 7th Sep., ie. restriction digestion with EcoR1 and the electrophoresis of the same. Sushma and Binu did the plasmid extraction with the cultures that was prepared yesterday (from MIT), for backup. The results of the gel electrophoresis was disappointing, again, as we didnt get the bands of the right size. But this gel also showed that the problem couldn't lie with the plasmids alone, as even pBluescript showed some bands of the size that was not expected. We along with Axel came to the conclusion that it probably was not just the plasmids but also the restriction enzyme, EcoR1 that was messing up the experiment. There was a slight possiblity that even RNAse could be the culprit. So,it was decided that on Tuesday, we will be digesting the plasmids (the third set and the one that was extracted today)with 3 restriction enzymes - new EcoR1, and the enzymes that would help to insert the gene into the plasmid, ie. Spe1 and Xba1. | ||
Tiffany ran the Formeldehyde Agarose Gel Electrophoresis on the ectracted RNA again today. The results again were that there was no RNA in the sample. This could possibly be due to not using RNAase free materials and equipment during the extraction process. We will discuss these results with Axel on Tuesday to determine our next step. | |||
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Revision as of 19:14, 9 October 2008
 A Flower of a Different Color by "Emblazon"
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Electrophoresis DayA Flower of A Different Color Today, like any other lab day, Angie, Diwash, Sushma and Binu, again, were working on the plasmid extraction hoping to get good results. Angie and Diwash worked on the plasmids that were extracted on 7th Sep., ie. restriction digestion with EcoR1 and the electrophoresis of the same. Sushma and Binu did the plasmid extraction with the cultures that was prepared yesterday (from MIT), for backup. The results of the gel electrophoresis was disappointing, again, as we didnt get the bands of the right size. But this gel also showed that the problem couldn't lie with the plasmids alone, as even pBluescript showed some bands of the size that was not expected. We along with Axel came to the conclusion that it probably was not just the plasmids but also the restriction enzyme, EcoR1 that was messing up the experiment. There was a slight possiblity that even RNAse could be the culprit. So,it was decided that on Tuesday, we will be digesting the plasmids (the third set and the one that was extracted today)with 3 restriction enzymes - new EcoR1, and the enzymes that would help to insert the gene into the plasmid, ie. Spe1 and Xba1. Tiffany ran the Formeldehyde Agarose Gel Electrophoresis on the ectracted RNA again today. The results again were that there was no RNA in the sample. This could possibly be due to not using RNAase free materials and equipment during the extraction process. We will discuss these results with Axel on Tuesday to determine our next step. | |
