User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/07/05

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] AU CHEM-570 Lab Prep
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 * style="background-color: #F2F2F2" align="center"|  |Main project page


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Objective
Add today's objective(s) here...

Bench work

 * 1) Analytical gel
 * 2) ladder
 * 3) Matt Hartings' sample
 * 4) - 6. --
 * 5) 5μL 1:10 pTXB1His
 * 6) 2.5μL gel-purified double-digested insert from  last week
 * 7) 2.5μL gel-purified double-digested vector(His) from  last week
 * 8) 2.5μL gel-purified double-digested vector from  last week
 * 9) 5μL V(His)+I ligation from  last week
 * 10) 5μL V+I ligation from  last week
 * 11) &rarr; 50' @ 90V
 * 12) &rarr; stain in EtBr
 * 13) Transformation
 * 14) 100μL DH10B + 5μL V(His)+I from  last week (same as what is on gel)
 * 15) 100μL DH10B + 5μL V+I from  last week (same as what is on gel)
 * 16) 100μL DH10B + 5μL V(His) neg ctrl from  last week
 * 17) 100μL DH10B + 5μL V neg ctrl from  last week
 * 18) * heat shock is 45s @ 42°C
 * 19) * plate 100μL on LBAmp100
 * 20) *&rarr; 37°C O/N
 * 21) Miniprep
 * 22) 10 mL LB + 10μL Amp100 + V+I in DH10B colony from  last week's transformation (100μL plate)
 * 23) 10 mL LB + 10μL Amp100 + V+I in NovaBlue colony from  last week's transformation, but the  450μL re-plate
 * 24) *&rarr; 37°C O/N w/ shaking
 * 1) *&rarr; 37°C O/N w/ shaking

Results

 * can't see undigested vector
 * gel-purified samples did not float away and are visible on gel (good!)
 * clearly see insert and double-inserts (~3kb). Faint band visible at correct size for vector+insert ligation (~7kb) but hard to see in photo.


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