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Things we did today

Wetlab work

PCR

Grace

• Colony PCR of nir-1, nir-2, GFPf-1, GFPf-2

• Annealed at 62C; extended for 1 min.
• Used all new PCR water, primer solution, and Green Taq
• Cut out bands for nir-1, GFP-1 (correct size)

• PCR of PCR components to test for contamination

• New primer solution + new PCR water + old Taq = no bands
• Old primer solution + new PCR water + new Taq = band at 300bp = primer solution was contaminated
• New primer solution + old PCR water + new Taq = bands at 270bp and 350bp = PCR water was contaminated

EtBr stained 2.5% agarose gel ran at 60V for 2 hours. Five microliters of each PCR reaction was loaded into each well.

Gel after excision of nir-1 and GFPf-1 bands.

EtBr stained 4% agarose gel ran at 95V for 90 min. Ten microliters of each PCR reaction was loaded into each well.

Cryostocked

Grace

• nir-1, GFPf-1

• Threw out old (incorrect) cryostocks of nir and GFPf

Discussion

Quote of the Day

History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson

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