User:Lu Wang/Notebook/Team Allergy/2010/07/06
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ProceduresToday, we isolated plasmid that contained antisense and sense sequences of the allergen panel and introns PME and PAL that we grew last week. For amiRNA
For ihpRNA
ResultsFor amiRNAMiniprep of turbo bacteria for plasmids containing sequences for amiRNA Should obtain plasmids containing the complete sequence for amiRNA. The resulting concentration should be relatively high (<100ng/μL) because all growing bacteria would have the plasmids inside. Nanodrop of miniprepped amiRNA plasmids We used 2μL of DNA per nanodrop.
Diagnostic digest of amiRNA plasmids Digested 200~500ng of plasmids with EcoR1 and Spe1. We will then run the digested vectors on an E Gel and look for DNA that is the length of the insert. If such DNA appears on the gel, then we will send it off to be sequenced. E Gel of digested amiRNA We had to run 21 samples, so we used two gels. The orders were: Gel #1
Gel #2
For ihpRNAMiniprep of plasmids containing introns for hpRNA Nanodrop of miniprepped intron plasmids We used 2μL of DNA per nanodrop.
Gel #1 Lane 1: 1kb Ladder Lanes 2-4: Ger Sense Lanes 5-7: Ger Antisense Lanes 8-10: LTP Sense Lanes 11-13: LTP Antisense Lanes 14-16: Bet 2 Sense Lanes 17-19: Bet 2 Antisense Lanes 20-22: Bet 1 Sense Lanes 23-25: Bet 1 Antisense
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