840:153g:Projects/project10/2010/10/14

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  • On Tuesday we looked at results of gel from PCR testing our primers. Results showed combinations 3 (forward with biobrick Sneezy + reverse without biobrick Cowboy) and 4 (forward without biobrick Grumpy + reverse with biobrick Cowgirl) worked the best. Image of gel is unavailable to upload - see lab books.

Also, we figured out the materials and volumes of materials to have ready for RNA extraction for Thursday. Prepared and carried out RNA extraction from liquid mycelium culture. Then, prepared and carried out Reverse Transcriptase PCR for primer matches 1 (forward with biobrick Sneezy + reverse with biobrick Cowgirl) and 4 (forward without biobrick Grumpy + reverse with biobrick Cowgirl). Will remove and store in freezer Friday morning. Next time will proceed based on the results of PCR. If primer 1 combination works, we will further amplify and put in plasmid PSB1C3. If we have to use combination 4, we will put into T vector plasmid and attempt TA cloning.

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