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From 2012.igem.org

Contents 1 I. Assay Continued 2 II. Went to pick up a PCR machine and a sonicating water bath 3 III. Measured primer DNA concentration 4 IV. Had meeting with Prof. Isaacs 5 V. Primer concentration and proportions for DNA solutions calculations:

I. Assay Continued

7. 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

8. Plates were set in 37C incubator overnight. The remaining cell-solutions were refrozen at -80C.

Andriana Notebook 2.jpg

Andriana Notebook 3.jpg

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