840:153g:Projects/project1/2008/11/06

A Flower of a Different Color by "Emblazon"

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Entry title A Flower of A Different Color We, vector group (Angie and Diwash) did the PCR of plasmids pSB1A3 and pSB1A7 by the use of forward and reverse primers. The primers should give 316 basepair fragments in the case of pSB1A3 and 590 basepairs in pSB1A7. If the fragments from PCR matches with the fragments of respective plasmids then they are not mislabeled, otherwise if the sizes are just opposite to one-another it can prove that the MIT samples are mislabeled. We have to wait for the result till next laboratory day (11th Nov. 08'). From the result we can also varify that whether our band sizes of plasmids pSB1A3 and pSB1A7 of the previous experiment (4th Nov. 8') are correct or not. Sushma and Binu performed a double digest of extracted plasmids pSB1A2, each with BBa_R0010, BBa_R0040, BBa_R0051 promoter to verify the promoters and plasmid identity. We used fast-digest enzymes, SpeI & XbaI to digest the plasmid DNA and then ran gel for 40 minutes at 150V. We got a band of 2000bp showing the identity of plasmid pSB1A2 and a band of 200bp for BBa_R0010.we also got a band of 2000bp for BBa_R0040 but the length for this promoter is 50bp which is too small .We dint get any band for BBa_R0051. Melissa, Tiffnay and Rajiv performed electrophoresis of the PCR DNA products from the leaf and the RNA extracted from the flower(both from the phalanopsis orchid plant).Tiffany had cleaned the RNA electrophoresis apparatus propr to the class. The DNA electrophoresis showed no amplification of the DNA of the PCR products, our positive control did not amplify either,from which we infer that it may be due to a problem with the PCR pre-,mix used during PCR ,it may be also due less concentration of the DNA in our samples.The other team (wintergreen) also used the same PCR pre-mix thier results will be compared during the next class if they have encountered a similar problem s ours. The RNA electrophoresis yielded positive results there were RNA bands seen against the RNA ladder used .Reverse PCR of the rna will be done during our next class to a DNA product ,this can be done only if the PCR issue is resolved.