Editing 1710-pT7A3-Nde1 (section)

=== '''Nde1 QC'''  ===

'''[[Image:Nde1 QC 7-24-2013.JPG]]''' 

enzyme dilution buffer: 20 mM Tris-HCl, 50 mM NaCl, 0.1 mM EDTA, 50% Glycerol, pH 7.5<br> 

{| width="447" border="1" cellspacing="1" cellpadding="1"
|-
| lane 
| 1035,pET28b-Tth-Ligase-K294R,200ng<br> 
| concentration(mg/ml)<br> 
| digestion time<br> 
| company<br>
|-
| 1 
| Nde1+BamH1<br> 
| 0.4<br> 
| 1 h<br> 
| Excellgen<br>
|-
| 2 
| Nde1+BamH1<br> 
| 0.6<br> 
| 1 h<br> 
| Excellgen<br>
|-
| 3 
| Nde1+BamH1<br> 
| 0.8<br> 
| 1 h<br> 
| Excellgen<br>
|-
| 4 
| Nde1+BamH1<br> 
| 1.2<br> 
| 1 h<br> 
| Excellgen<br>
|-
| 5 
| Nde1+BamH1<br> 
| 1.5<br> 
| 1 h<br> 
| Excellgen<br>
|-
| 6 
| Nde1+BamH1<br> 
| <br> 
| 1 h<br> 
| NEB<br>
|}

<br>

9/29/2013<br>

Purification of Nde1 using 25 ml heparin HyperD column (XK16).<br>

Dilute partial purified Nde1 with 50 mM Tris-HCl. Load to heparin column.<br>

Elute with 250 ml linear gradient of 0.1 M to 0.6 M NaCl. Nde1 is eluted at 0.4 M NaCl.<br>

<br>

[[Image:Nde1_purification.jpg]]<br> 

<br>

{| width="305" height="336" border="1" cellspacing="1" cellpadding="1"
|-
| Lane<br>
| Clone ID<br>
| Desc<br>
|-
| 1<br>
| 1710<br>
| Nde1, Eluted from Heparin column<br>
|-
| 2<br>
| 1710<br>
| Nde1, Eluted from Heparin column<br>
|-
| 3<br>
| 1710<br>
| Nde1, Eluted from Heparin column<br>
|-
| 4<br>
| 1813<br>
| FLP, 50 mM I wash from Ni column <br>
|-
| 5<br>
| 1813<br>
| FLP, 300 mM I eluted from Ni, diluted<br>
|-
| 6<br>
| 1710<br>
| Nde1, Eluted from Heparin column<br>
|-
| 7<br>
| 1813<br>
| FLP, 300 mM I eluted from Ni, diluted<br>
|-
| 8<br>
| 1813<br>
| FLP, heparin column FT<br>
|-
| 9<br>
| 1813<br>
| FLP, 50 mM I wash from Ni column<br>
|-
| 10<br>
| 1835<br>
| MBP-Taq-Muts-6XHis, partial purified from Ni column<br>
|-
| 11<br>
| 1813<br>
| FLP heparin column elute<br>
|-
| M<br>
| <br>
| Fermentas PageRuler Plus<br>
|-
| 12<br>
| 1813<br>
| FLP heparin column elute<br>
|-
| 13<br>
| 1813<br>
| FLP heparin column elute<br>
|-
| <br>
| <br>
| <br>
|}

<br>