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MS reports 5 30 2017-6 2 2017 - Revision history
2026-10-18T02:39:36Z
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Crm at 20:59, 2 June 2017
2017-06-02T20:59:39Z
<p></p>
<p><b>New page</b></p><div>== 05/30/2017 ==<br />
<br />
Culture 1782 <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; orphans: 2; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; -webkit-text-stroke-width: 0px; background-color: rgb(255, 255, 255); text-decoration-style: initial; text-decoration-color: initial; display: inline !important; float: none">Tth Ligase (8L), induced with IPTG, over night.</span><br />
<br />
<span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; orphans: 2; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; -webkit-text-stroke-width: 0px; background-color: rgb(255, 255, 255); text-decoration-style: initial; text-decoration-color: initial; display: inline !important; float: none">Culture 483 Tth Polymerase (200ml), over night.</span><br />
<br />
<span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; orphans: 2; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; -webkit-text-stroke-width: 0px; background-color: rgb(255, 255, 255); text-decoration-style: initial; text-decoration-color: initial; display: inline !important; float: none">Prepare Ni Elution Buffer (50mM phosphate buffer, pH7.0, 250mM NaCl, 100mM Arginine, 300mM imidazole, 10% glycerol)</span><br />
<br />
<span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; orphans: 2; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; -webkit-text-stroke-width: 0px; background-color: rgb(255, 255, 255); text-decoration-style: initial; text-decoration-color: initial; display: inline !important; float: none">Wash Ni column (bind with 1708 HTN Cre) with Ni washing Buffer, phosphate buffer and Ni Elution buffer.</span><br />
<br />
<span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; orphans: 2; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; -webkit-text-stroke-width: 0px; background-color: rgb(255, 255, 255); text-decoration-style: initial; text-decoration-color: initial; display: inline !important; float: none">Prepare SDS-PAGE Gel Stain.</span><br />
<br />
<span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; orphans: 2; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; -webkit-text-stroke-width: 0px; background-color: rgb(255, 255, 255); text-decoration-style: initial; text-decoration-color: initial; display: inline !important; float: none">Prepare Buffer A2, SP Dilution Buffer, Ni Elution Buffer.</span><br />
<br />
== 05/31/2017 ==<br />
<br />
Culture 483 Tth Polymerase (8L), induced with IPTG.<br />
<br />
Spin down 1782 Tth ligase, homogenize, centrifugation, load onto Ni column.<br />
<br />
Prepare LB media.<br />
<br />
Prepare SP Washing Buffer and SP Elution Buffer.<br />
<br />
Elute SP FF column (bind with 1708 HTN Cre) with SP washing buffer and SP Elution buffer. Dialysis over night.<br />
<br />
[[File:123-1708 HTN Cre SP FF 5 31 2017.jpg|RTENOTITLE]]<br />
<br />
Culture 1708 HTN Cre (200ml).<br />
<br />
== 06/01/2017 ==<br />
<br />
Culture 1708 HTN Cre (8L), induced with IPTG, over night.<br />
<br />
Spin down 483 Tth Polymerase.<br />
<br />
Dialysis of 1708 HTN Cre, total 37ml, 1.07mg/ml. Need to further dialyse.<br />
<br />
Culture 1464 T7 DNA ligase (200ml)<br />
<br />
Wash Ni column (bind with 1782 Tth ligase) with Ni washing Buffer/ Tris-HCl/ Buffer B,<br />
<br />
Elute 1782 Tth ligase with Buffer B 06/01/2017<br />
<br />
{| style="width: 500px" cellspacing="1" cellpadding="1" border="1"<br />
|-<br />
| -0.08<br/><br />
| 0.12<br/><br />
| 1.07<br/><br />
| 3.23<br/><br />
| 5.68<br/><br />
| (1~5)<br/><br />
|-<br />
| 3.70<br/><br />
| 2.49<br/><br />
| 1.46<br/><br />
| 0.87<br/><br />
| 0.59<br/><br />
| (6~10)<br/><br />
|-<br />
| 0.43<br/><br />
| 0.31<br/><br />
| 0.30<br/><br />
| 0.19<br/><br />
| 0.11<br/><br />
| (11~15)<br/><br />
|}<br />
<br />
load sample 3~12 onto QHP column.<br />
<br />
== 06/02/2017 ==<br />
<br />
Regenerate Ni column and SP FF column.<br />
<br />
Elute 1782 Tth Ligase.<br />
<br />
[[File:124-1782 Tth Ligase QHP 6 2 2017.jpg]]<br />
<br />
Homogenize 483 Tth Polymerase, spin down, load supernatant onto Ni column.<br />
<br />
Deal with order 4874, ship to Neitherlands.<br />
[[Category:Reports]]</div>
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