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MS 6 9 2017 - Revision history
2026-10-18T10:23:35Z
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Crm at 15:15, 12 June 2017
2017-06-12T15:15:48Z
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<p><b>New page</b></p><div>1518 T3 Ligase, 1601 6His-GST-i3C, 1765 T4 DNA Polymerase, 1942 phi 29 pick up single colony to culture in LB/K/C media for small scale screening.<br />
<br />
Spin down 1518 T3 Ligase, 1601 6His-GST-i3C, 1765 T4 DNA Polymerase, 1942 phi 29 to make glycerol stock. Store at -80<sup>o</sup>C.<br />
<br />
Spin down 1498 Taq DNA Ligase, store at -20<sup>o</sup>C.<br />
<br />
Run SDS-PAGE gel of 1464&nbsp;1464 T7 DNA Ligase 06/09<br />
<br />
{| style="width:500px" cellspacing="1" cellpadding="0" border="1" width="500"<br />
|-<br />
| <br />
Number<br />
<br />
| <br />
1<br />
<br />
| <br />
2<br />
<br />
| <br />
3<br />
<br />
| <br />
4<br />
<br />
| <br />
5<br />
<br />
| <br />
6<br />
<br />
| <br />
7<br />
<br />
| <br />
8<br />
<br />
| <br />
9<br />
<br />
|-<br />
| <br />
sample<br />
<br />
| <br />
13<br />
<br />
| <br />
14<br />
<br />
| <br />
15<br />
<br />
| <br />
16<br />
<br />
| <br />
17<br />
<br />
| <br />
18<br />
<br />
| <br />
19<br />
<br />
| <br />
20<br />
<br />
| <br />
MW<br />
<br />
|-<br />
| <br />
uL<br />
<br />
| <br />
9.1<br />
<br />
| <br />
3.1<br />
<br />
| <br />
0.8<br />
<br />
| <br />
0.4<br />
<br />
| <br />
0.3<br />
<br />
| <br />
0.9<br />
<br />
| <br />
1.5<br />
<br />
| <br />
3.7<br />
<br />
| <br />
10<br />
<br />
|-<br />
| <br />
ddH<sub>2</sub>O<br />
<br />
| <br />
5<br />
<br />
| <br />
10<br />
<br />
| <br />
15<br />
<br />
| <br />
15<br />
<br />
| <br />
15<br />
<br />
| <br />
15<br />
<br />
| <br />
15<br />
<br />
| <br />
10<br />
<br />
| <br />
\<br />
<br />
|}<br />
<br />
[[File:Protein Expression SDS-PAGE 126-1464 T7 DNA Ligase QHP 6 9 2017.jpg|RTENOTITLE]]<br />
<br />
Collect 4~8 forward to the next step. Dialysis.<br />
<br />
23.2mg/ml, 4.5ml, 104.4mg. Purity>95%.<br />
<br />
Wash Ni column (load with 1946 TdT bovine) with Ni washing buffer / Tris-HCl / Buffer B.<br />
<br />
Elute 1946 TdT bovine with Buffer B 06/09/2017<br />
<br />
{| style="width: 500px" cellspacing="1" cellpadding="1" border="1"<br />
|-<br />
| -0.05<br/><br />
| -0.21<br/><br />
| -0.07<br/><br />
| 0.81<br/><br />
| 2.81<br/><br />
| (1~5)<br/><br />
|-<br />
| 2.05<br/><br />
| 1.09<br/><br />
| 0.71<br/><br />
| 0.50<br/><br />
| 0.33<br/><br />
| (6~10)<br/><br />
|-<br />
| 0.23<br/><br />
| 0.18<br/><br />
| 0.13<br/><br />
| 0.05<br/><br />
| 0.01<br/><br />
| (11~15)<br/><br />
|}<br />
<br />
collect 4~12<br />
[[Category:My ELN]]</div>
Crm