From 2008.igem.org

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Things we did today

Wetlab work

Construction of p+r and re-replacement of BB-pRL1383a MCS

Grace

EtBr stained 2% (L) and 0.8% (R) agarose gels ran at 60V for 2 hours and 1 hour, respectively. Thirty microliters of RE digest reaction were loaded into each well.

• Ran RE digests from last night on agarose gel

• C0012: correct vector band ~2.1kb
• J33207: band at ~700bp (only one enzyme cut -- XbaI did not cut; PstI cuts because C0012 was cut by PstI) and ~850bp (uncut PCR product)
• nir: ~800bp band (from GFP contaminant in plasmid prep, cut only once) and ~2kb band (??)
• BB-pRL1383a: ran WAY too much product. Will redo RE digest with 20% of plasmid used. Since J33207 was only cut once, it's assumed BB-pRL1383a was cut only once as well.

• Extracted C0012 vector band from gel
• Ligated:

• plac + rbs (B0034)

• PCR of J33207
• RE digested:

• p+r with EcoRI, XbaI, SpeI, PstI
• J33207 and BB-pRL1383a with EcoRI and PstI

• Treated C0012 vector (pSB1A2) with SAP

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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