Ruili.su at 18:59, 15 September 2011

2011-09-15T18:59:41Z

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= PEG-Plasmid-Max-Prep(compare PEG/NaCl/MgCl2 with PEG/CaCl2) =

Reference to the protocol: [[PEG-Plasmid-Max-Prep|PEG-Plasmid-Max-Prep]]. 

Grow STB3 transformed with clone 303 in 2 flasks (500 ml) of LB/AMP. Precipitate with CTAB, then dissolve DNA in 0.75M NaCl, precipitate with PEG/NaCl/MgCl2 or PEG/CaCl2

I.Purity by OD260/280 

{| cellspacing="2" cellpadding="1" border="1" style="width: 389px; height: 122px;"
|-
| Sample 
| OD260/280 
|   Concentration 
|-
| 1. PEG/NaCl/MgCl2 
|   1.89 
|   1474.5ng/ul 
|-
| 2. PEG/NaCl/MgCl2 
|   1.90 
|   762.5ng/ul 
|-
| 1. PEG/CaCl2 
|   1.87 
|   962.8ng/ul 
|-
| 2. PEG/CaCl2 
|  1.83 
|   237.7ng/ul 
|}

 

1----the first time to dissolve DNA with water;

2----the second time to dissolve DNA with water;

II.Run Argrose Gel.

[[Image:PEG-Plasmid-Maxi-Prep.JPG|243x345px|PEG-Plasmid-Maxi-Prep.JPG]] 

1 and 2 are the plasmid extracted by PEG/CaCl2, which does not have RNA; 

3 and 4 are the plasmid extracted by PEG/NaCl/ MgCl2, which has the trace of RNA;

According to I and II, PEG/CaCl2 is good. So we can make sure that using PEG/CaCl2 to extract DNA for Maxi Prep should be better.

 

--[[User:Ruili.su|Ruili.su]] 14:51, 14 September 2011 (EST)

[[Category:Cloning]]

PEG-Plasmid-Max-Prep Precipitation - Revision history

Ruili.su at 18:59, 15 September 2011