Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2015/02/12

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I cut some of the PCR product I created on 2/5/2015 with PstI and BbsI again (see protocol on 2/6/2015).
This time, I completed the protocol 4 times in 4 separate tubes. Then, I PCR purified the 4 tubes all in the same column. (20μL in each tube = 80μL total for PCR purification. During the elution step, I eluted my column with 20μL of elution solution instead of 50μL to increase my final concentration. After the first round of elution, I kept the column and applied another 20μL of elution solution to it, and eluted again. The first elution is expected to have a significantly higher concentration than the second.

PCR Purification results

Sample Read# 260 280 260/280 ng/µL
1 (first elution) 0.062 0.036 1.746 62.034
2 (second elution) 0.013 0.01 1.391 13.462