Editing Plasmid-CTAB-Max-Prep (section)

== Buffers<br>  ==

'''TE (Tris/EDTA)''' [500 mL]<br>25 mM Trisâ¢HCl pH 8.0 [6.25 mL 2M]<br>10 mM EDTA [10 mL 0.5M]<br> 

Autoclave 

Add lysozyme<br> 

<br>'''P3'''<br> 

3.0 M potassium acetate pH 5.5<br>Dissolve 294.5 g potassium acetate in 500 ml dH2O. Adjust the pH to 5.5 with glacial acetic acid (~110 ml). Adjust the volume to 1 liter with dH2O. 

for 500 ml: 147g KOAc (3M)<br>55 mL glacial HOAc<br> 

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'''P3U buffer''' 

3M KAc, 5M CH3COOH, 4M Urea.<br> 

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'''PEG/NaCl/MgCl<sub>2</sub> solution'''<br>30% w/v PEG 8000<br>1.6M NaCl<br>25 mM MgCl<sub>2</sub>. 

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'''PEG-MgCl'''<sub>'''2'''</sub><br>40% (w/v) polyethylene glycol (PEG 8000) 

30 mM MgCl<sub>2</sub> 

Dissolve 40 g of PEG 8000 in a final volume of 100 ml of 30 mM MgCl2. Sterilize the solution by passing it through a 0.22-µm filter, and store it at room temperature.<br> 

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'''Ca/PEG'''<br>0.4 M calcium chloride<br>40% PEG 8000<br> 

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'''CTAB/NaCl'''<br> 

2% (w/v) CTAB<br> 

0.4 M NaCl<br> 

'''<br>2% (w/v) CTAB'''<br> 

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Note: it was reported that RNA can be precipitated with 1.4M CaCl<sub>2</sub> (add with P3), then plasmid DNA can be precipiated with 10% polyethylene glycol (PEG 8000), 250 mM NaCl. http://www.ncbi.nlm.nih.gov/pubmed/18571491<br> 

[[Category:Protocols]]