User:Robwarden/Notebook/Divalent Ligand Validation/2009/03/14

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 * style="background-color: #EEE"|[[Image:TBL.PNG|300px]] Divalent Ligand Validation
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 * style="background-color: #F2F2F2" align="center"|  |Main project page


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Make New Amp Plates

 * Amp100
 * Spit Test

Reactions

 * pMSCVN (XhoI & NotI) + H1C (XhoI & NotI)
 * pMSCVN (XhoI & NotI) + H1Y (XhoI & NotI)
 * pMSCVN (XhoI & NotI) + Water (Negative Control)
 * DNA from 25 Feb 09

Setup
9.4 μL H1C/H1Y/Water digest 0.6 μL pMSCVN digest 10 μL Quick Ligation Buffer 1 μL Quick Ligase

Conditions
Incubate 5 min at room temperature.

Reaction
50 μL XL10-Blue Supercompetent Cells 0.85 μL beta-mercaptoethanol 5 μL Ligation Product

Conditions

 * 1) Ice for 30 min.
 * 2) 42°C Water bath for 45 sec.
 * 3) Ice for 2 min.  Add 450μL preheated SOC.
 * 4) Shake at 37°C for 1 hr.
 * 5) Plate 150mL on each plate.
 * 6) Incubate at 37°C overnight.


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