Crm at 19:53, 7 July 2017

2017-07-07T19:53:35Z

New page

== 07/03/2017 ==

Culture 1567 Exo 200ml. Culture Tn5 transposase 8L, induced with IPTG.

Homogenize 1811 vPol, load onto Ni column.

Load 1458 KOD onto QHP column.

Ship order 4890 and 4891.

Prepare SDS-PAGE Gel staining.

== 07/04/2017 ==

Culture 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">pET30b-DeepVent-sso7d 200ml. Culture 1567 Exo 8L, induced with IPTG.

Homogenize 2462 Tn5 transposase, load onto Ni column.

Load 1995 Kfu onto QHP column.

Wash Ni column (load with 1811 vPol) with Ni washing buffer/ Tris-HCl/ Buffer B.

Elute 1811 vPol with Buffer B 07/04/2017

{| style="width: 500px" cellspacing="1" cellpadding="1" border="1"
|-
| -0.02 
| -0.11 
| -0.14 
| -0.21 
| -0.08 
| (1~5) 
|-
| 1.18 
| 3.41 
| 6.35 
| 17.97 
| 36.35 
| (6~10) 
|-
| 20.81 
| 15.44 
| 10.71 
| 4.83 
| 1.61 
| (11~15) 
|-
| 0.76 
| 0.32 
| 0.24 
| 0.12 
| 0.08 
| (16~20) 
|}

collect 6~17.

Elute 1458 KOD

[[File:136-1458 KOD QHP 7 4 2017.jpg|RTENOTITLE]]

Run SDS-PAGE gel of 1458 KOD  7.91/92.2   07/04/2017

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

|
1

|
2

|
3

|
4

|
5

|
6

|
7

|
8

|
9

|
10

|
11

|
12

|-
|
sample

|
16

|
18

|
20

|
22

|
24

|
26

|
28

|
30

|
MW

|
32

|
34

|
36

|-
|
uL

|
10

|
0.2

|
0.1

|
0.13

|
0.13

|
0.2

|
0.23

|
0.7

|
10

|
2.0

|
4.2

|
7.7

|-
|
ddH2O

|
5

|
15

|
15

|
15

|
15

|
15

|
15

|
15

|
\

|
15

|
10

|
5

|}

[[File:Protein Expression SDS-PAGE 136-1458 KOD QHP 7 4 2017.jpg|RTENOTITLE]]

forward lane 1~3 to the next step. 

== 07/05/2017 ==

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 200ml. Culture 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">pET30b-DeepVent-sso7d 8L, induced with IPTG.

Homogenize 1567 Exo, load onto Ni column.

Load 1811 vPol onto QHP column.

Wash Ni column (load with 2462 Tn5 transposase) with Ni washing buffer/ Tris-HCl/ Buffer B.

Elute 2462 Tn5 transposase with Buffer B 07/05/2017

{| style="width: 500px" cellspacing="1" cellpadding="1" border="1"
|-
| -0.24 
| -0.26 
| -0.24 
| -0.23 
| -0.20 
| (6~10) 
|-
| -0.15 
| 0.28 
| 1.31 
| 0.38 
| 0.04 
| (11~15) 
|-
| 0.00 
| 
| 
| 
| 
| (16~20) 
|}

collect 12~15.

Elute 1995 Kfu

[[File:137-1995 Kfu QHP 7 5 2017.jpg|RTENOTITLE]]

Run SDS-PAGE gel of 1995 Kfu 7.87 / 99.3     07/05/2017

{| 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

|
17

|
18

|
19

|
20

| style="width:33px" |
21

| style="width:33px" |
22

|
23

|
MW

|
25

|
27

|
29

|
31

|-
|
uL

|
9.1

|
1.5

|
0.8

|
0.7

| style="width:33px" |
1.1

| style="width:33px" |
1.7

|
2.4

|
10

|
3.3

|
4.4

|
6.7

|
11.1

|-
|
ddH2O

|
5

|
15

|
15

|
15

| style="width:33px" |
15

| style="width:33px" |
15

|
15

|
\

|
10

|
10

|
10

|
5

|}

[[File:Protein Expression SDS-PAGE 137-1995 Kfu QHP 7 5 2017.jpg|RTENOTITLE]]

forward lane 1~4 to the next step.  

Dialysis of 1458 KOD. 14.27mg/ml, 4.5ml, purity > 90%

== 07/06/2017 ==

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.

Homogenize 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">pET30b-DeepVent-sso7d, load onto Ni column.

Load 2462 Tn5 transposase onto QHP column.

Wash Ni column (load with 1567 Exo) with Ni washing buffer/ Tris-HCl/ Buffer B.

Elute 1567 Exo with Buffer B 07/06/2017

{| style="width: 500px" cellspacing="1" cellpadding="1" border="1"
|-
| -0.25 
| -0.26 
| -0.25 
| -0.25 
| -0.23 
| (1~5) 
|-
| -0.10 
| 1.82 
| 1.04 
| 0.36 
| 0.15 
| (6~10) 
|-
| 0.07 
| 0.04 
| 0.03 
| 0.01 
| 0.01 
| (11~15) 
|}

collect 7~11.

Elute 1811 vPol

[[File:138-1811 vPol QHP 7 6 2017.jpg|RTENOTITLE]]

Run SDS-PAGE gel of 1811 vPol 6.63 / 69.2     07/06/2017

{| 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

|
8

|
10

|
12

|
14

| style="width:33px" |
15

| style="width:33px" |
16

|
17

|
MW

|
18

|
19

|
20

|
21

|-
|
uL

|
5.9

|
3.9

|
4.0

|
2.1

| style="width:33px" |
0.23

| style="width:33px" |
0.38

|
0.3

|
10

|
0.44

|
0.55

|
0.7

|
0.1

|-
|
ddH2O

|
10

|
10

|
10

|
15

| style="width:33px" |
15

| style="width:33px" |
15

|
15

|
\

|
15

|
15

|
15

|
15

|}

[[File:Protein Expression SDS-PAGE 138-1811 vPol QHP 7 6 2017.jpg|RTENOTITLE]]

forward lane 1~7 to the next step.  

Dialysis of 1995 Kfu. 4.18mg/ml, 4.5ml, purity > 90%

== 07/07/2017 ==

Wash Ni column (load with 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) with Ni washing buffer/ Tris-HCl/ Buffer B.

Elute 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 with Buffer B 07/07/2017

{| style="width: 500px" cellspacing="1" cellpadding="1" border="1"
|-
| -0.29 
| -0.26 
| -0.14 
| 0.99 
| 1.27 
| (6~10) 
|-
| 0.94 
| 0.80 
| 0.70 
| 0.64 
| 0.61 
| (11~15) 
|-
| 0.57 
| 0.50 
| 0.49 
| 0.44 
| 0.40 
| (16~20) 
|-
| 0.39 
| 0.37 
| 0.30 
| 0.28 
| 0.24 
| (21~25) 
|-
| 0.22 
| 0.22 
| 0.16 
| 0.11 
| 0.08 
| (26~30) 
|}

collect 9~27.

Elute 2462 Tn5 transposase

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

Run SDS-PAGE gel of 2462 Tn5 transposase  8.41 / 83.8    07/07/2017

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

|
1

|
2

| style="width:33px" |
3

| style="width:33px" |
4

|
5

|
6

|
7

|
8

|
9

|
10

|-
|
sample

|
22

|
23

| style="width:33px" |
24

| style="width:33px" |
25

|
26

|
MW

|
27

|
28

|
29

|
30

|-
|
uL

|
6.3

|
8.4

| style="width:33px" |
7.2

| style="width:33px" |
5.6

|
5.3

|
10

|
6.3

|
7.2

|
8.4

|
10

|-
|
ddH2O

|
10

|
5

| style="width:33px" |
5

| style="width:33px" |
10

|
10

|
\

|
10

|
5

|
5

|
5

|}

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

forward lane 1~4 to the next step.  

Dialysis of 1811 vPol. 36.0mg/ml, 10.0ml, purity > 90%
[[Category:Reports]]

MS reports 7 3 2017-7 7 2017 - Revision history

Crm at 19:53, 7 July 2017