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Vectors, RNA, and PCR
Our Field of Dreams
The Gene group (Rajiv, Melissa, and Tiffany) performed an RNA extraction using a RNAeasy Mini kit provided by Axel. An electrophoresis will be performed next Tuesday to see if RNA was actually extracted. A mistake was made during the extraction and the waste from the RNA was placed into the tube containing the DNA PCR product. So the PCR was repeated with 5 tubes of each of the extracted DNA samples. The results of the PCR will be analyzed using electrophoresis next Tuesday as well.
The vector group(Angie,Binu,Sushma,Diwash)performed three restriction digests for each plasmid sample using a different enzyme for each digest as we did on tuesday. The enzymes used were fast digest Spe1,normal EcoR1,and fast digest Xbal. As we saw the samples of pSB1A3 and pSB1A7 from Dr Axel’s lab din’t worked we decided to work only with glycerol stocks that we got from MIT . We ran the electrophoresis of our digest plasmids and also of undigested plasmids to make sure our plasmids were present in the sample. We used the same protocol as we did on Tuesday but the bands were irregular .The bands which we got with the same restriction enzymes were not seen in todays gel.There is no problem with the gel because the ladder runned properly with all the bands. So we decided to extract plasmids again along with Dr.Axel and to verify the identity of these plasmids. (Binu and Sushma)We also decide to work on the promoter and to extract it from the partsregistery.