Team:Yale/Our Project/Notebook/Week 2

• home
• our project
• our team

our project

• introduction
• applications
• methods
• lab notebook
• week 1
• week 2
• week 3
• week 4
• week 5
• week 6
• week 7
• week 8
• protocols
• safety
• future work

lab notebook: week 2 (6/14-6/20)

• Monday 6/14--Redo of 6/11 assay of bacterial growth within a narrow copper(II) concentrations after analysis of data showed uniformly poor growth.
See more/less

Copper Growth Assay Work Continues

• As the growth assay of 6/11 6/11 showed uniformly poor growth even at the lowest copper concentrations (see above), redid it with care not to let the liquid cultures overgrow, a possible source of the cultures' previous poor performance. (add plot)
• In redoing plate assay, changed copper concentration line-up slightly, eliminating 1.25 mM and 2.5 mM trials to allow room for the control concentrations, 0 and 1 M, and only worked with high because there was no real distinction in the three different OD trials done on 6/10. For simplicity, future assays will only use one starting OD (0.075) per strain.
• During the repeat, noticed that DH5alpha liquid culture grown up for plate reader assay grew at a rate much slower than that of LE392, creating a need to keep diluting the LE392 solution with more LB so it would not overgrow before the DH5alpha was ready. In the end, the culture of LE392 used in the assay was diluted from an OD of 0.761 and the DH5alpha culture from an OD of 0.841.

This day's work is also recorded on pages 8-10 of the lab notebook hard copy.
• Tuesday 6/15--Did work on design of primers for PCR of thiosulfate reducates gene & Biobrick synthesis planning + set-up test to see if cultures from 6/14 survived the copper exposure.
See more/less
Extra content
• Wednesday 6/16--checked on spotted cell survival assay, collected MOPS minimal media materials, started making component solutions
See more/less
Extra content
• Thursday 6/17--more minimal media work and meeting, started BL21 culture
See more/less
Extra content
• Friday 6/18--plasmid pSB74 arrived (!), already in some unspecified E coli, so plated them out. Transformed promoters (constitutive & IPTG inducible), terminator, and repressor (for inducible promoter) into BL21. Also ran BL21 copper growth assays for wide and narrow concentrations ranges. June the 19th--in which it is revealed that there was a transformation-fail Redid transformation, this time into LE392, set aside pSB74 containing cells in fridge
See more/less
Extra content
• Sunday 6/20--in evening inoculate 5 mL liquid cultures
See more/less
Extra content