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User:Andriana/14 June 2012
From 2012.igem.org
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I. Assay Continued
- After being frozen for 24 hours, BL 21 cell samples (normal and transformed – normal on plain agar plates and transformed on amp+kan plates) were plated on agar plates
- Due to high probability of cell death during vortexing, an estimated 10,000 cells/plate were added to each plate by diluting 10 microliters from each of the eppendorf tubes in 200 microliters of DIW
- Plates were set in 37C incubator overnight. The remaining cell-solutions were refrozen at -80C.
Andriana Notebook 2.jpg
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Andriana Notebook 3.jpg
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II. Went to pick up a PCR machine and a sonicating water bath
III. Measured primer DNA concentration
IV. Had meeting with Prof. Isaacs
V. Primer concentration and proportions for DNA solutions calculations:
- [Micromolar] from spec: = [x from spectrophotometer] * 10^3/ molecular weight
- Making 25 micromolar solution so as to add to 1 microliter of primers to 50 microliters of a PCR reaction
- Amount of DIW for 1 microliter of primer stock = ([x concentration from spectrophotometer]*10^3) / (4*molecular weight) - 1
