Crm at 19:21, 14 July 2017

2017-07-14T19:21:07Z

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== 07/10/2017 ==

Culture 818 pHis-GST-His-Flag-His 8L, induced with IPTG.

Homogenize 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, centrifugate and load onto Ni column.

<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">Load 1567 Exo onto QHP column.

[http://excellgen.net/eln/index.php/MS_7_7_2017 <span style="font-family: sans-serif; font-size: 14px; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; 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">http://excellgen.net/eln/index.php/MS_7_7_2017]

[[File:139-2462 Tn5 transposase QHP 7 7 2017.jpg|RTENOTITLE]]

[[File:Protein Expression SDS-PAGE 139-2462 Tn5 transposase QHP 7 7 2017.jpg|RTENOTITLE]]

<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">Dialysis 2462 Tn5 Transposase. 1.48mg/ml, 5.5ml, purity >90%.

== 07/11/2017 ==

Homogenize 818 pHis-GST-His-Flag-His<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">, centrifugate and load onto Ni column.

<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 (load with 1782 Tth-Ligase) with Ni washing buffer/ Tris-HCl/ Buffer B. Elute <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">1782 Tth-Ligase with Buffer B 07/11/2017

{| style="width: 500px" cellspacing="1" cellpadding="1" border="1"
|-
| -0.26 
| -0.27 
| -0.28 
| -0.19 
| 0.03 
| (1~5) 
|-
| 4.68 
| 6.36 
| 7.35 
| 5.35 
| 4.74 
| (6~10) 
|-
| 4.15 
| 3.85 
| 3.53 
| 2.75 
| 2.35 
| (11~15) 
|-
| 1.85 
| 1.53 
| 1.21 
| 0.54 
| 0.23 
| (16~20) 
|-
| 0.12 
| 0.08 
| 
| 
| 
| (21~25) 
|}

collect 6~20.

<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">Load 423 DeepVent-sso7d onto QHP column.

Elute 1567 T5-Exo

[[File:140-1567 T5-Exo QHP 7 11 2017.jpg|RTENOTITLE]]

Run SDS-PAGE gel of 1567 T5-Exo 5.55 / 34.5    07/11/2017

{| style="width:500px" width="500" cellspacing="1" cellpadding="0" border="1"
|-
|
Number

|
1

|
2

|
3

|
4

| style="width:33px" |
5

| style="width:33px" |
6

|
7

|
8

|
9

|
10

|
11

|
12

|-
|
sample

|
19

|
20

|
21

|
22

| style="width:33px" |
23

| style="width:33px" |
24

|
25

|
MW

|
26

|
27

|
28

|
30

|-
|
uL

|
9.1

|
5.3

|
3.9

|
3.0

| style="width:33px" |
3.1

| style="width:33px" |
2.6

|
2.4

|
10

|
3.3

|
4.8

|
5.3

|
9.1

|-
|
ddH2O

|
5

|
10

|
10

|
10

| style="width:33px" |
10

| style="width:33px" |
10

|
10

|
\

|
10

|
10

|
10

|
5

|}

 [[File:Protein Expression SDS-PAGE 140-1567 T5-Exo QHP 7 11 2017.jpg|RTENOTITLE]]

Forward lane 11~12 to the next step.

== 07/12/2017 ==

<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 (load with 818 pHis-GST-His-Flag-His) with Ni washing Buffer/ Tris-HCl/ Buffer B

Elute 818 pHis-GST-His-Flag-His with Buffer B 07/12/2017

{| style="width: 500px" cellspacing="1" cellpadding="1" border="1"
|-
| -0.30 
| -0.26 
| 0.05 
| 1.95 
| 2.26 
| (6~10) 
|-
| 1.68 
| 1.18 
| 0.85 
| 0.62 
| 0.43 
| (11~15) 
|-
| 0.30 
| 0.20 
| 0.14 
| 0.08 
| 0.05 
| (16~20) 
|}

collect 9~16.

Load <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">1782 Tth-Ligase onto QHP column.

Elute <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">423 DeepVent-sso7d from QHP column.

[[File:141-423 DeepVent-sso7d QHP 7 12 2017.jpg|RTENOTITLE]]

<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">Run SDS-PAGE gel of 423 DeepVent-sso7d  8.69 / 99.5    07/12/2017

{| style="width:500px" width="500" cellspacing="1" cellpadding="0" border="1"
|-
|
Number

|
1

|
2

|
3

|
4

| style="width:33px" |
5

| style="width:33px" |
6

|
7

|
8

|-
|
sample

|
14

|
15

|
16

|
17

| style="width:33px" |
18

| style="width:33px" |
MW

|
20

|
22

|-
|
uL

|
15

|
4.6

|
2.7

|
2.7

| style="width:33px" |
4.0

| style="width:33px" |
10

|
5.0

|
5.9

|-
|
ddH2O

|
\

|
10

|
10

|
10

| style="width:33px" |
10

| style="width:33px" |
\

|
10

|
10

|}

 [[File:Protein Expression SDS-PAGE 141-423 DeepVent-sso7d QHP 7 12 2017.jpg|RTENOTITLE]]

Forward lane 1~2 to the next step.

Dialysis of 1567 T5-Exo. 1.5mg/ml, 8.0ml.

== 07/13/2017 ==

Load 818 pHis-GST-His-Flag-His <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">onto QHP column.

 Elute <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">1782 Tth-Ligase from QHP column.

[[File:142-1782 Tth ligase QHP 7 13 2017.jpg]]

<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">Run SDS-PAGE gel of 1782 Tth-Ligase   6.56 / 79.1    07/13/2017

{| style="width:500px" width="500" cellspacing="1" cellpadding="0" border="1"
|-
|
Number

|
1

|
2

|
3

|
4

| style="width:33px" |
5

| style="width:33px" |
6

|
7

|-
|
sample

|
MW

|
FT1

|
14

|
17

| style="width:33px" |
18

| style="width:33px" |
23

|
FT2

|-
|
uL

|
10

|
15

|
10

|
0.5

| style="width:33px" |
0.22

| style="width:33px" |
0.9

|
15

|-
|
ddH2O

|
\

|
\

|
5

|
15

| style="width:33px" |
15

| style="width:33px" |
15

|
\

|}

FT1: flow through after Ni column.

FT2: flow through after QHP column.

[[File:Protein Expression SDS-PAGE 142-1782 Tth ligase QHP 7 13 2017.jpg]]

Forward lane 5~6 to the next step.

Dialysis of 423 <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">DeepVent-sso7d. 1.50mg/ml, 8.0ml.

== 07/14/2017 ==

Dialysis of 1782 Tth-Ligase. 11.20mg/ml, 5.9ml.
[[Category:Reports]]

MS reports 7 10 2017-7 14 2017 - Revision history

Crm at 19:21, 14 July 2017