Leaking test
add H2O
-
00:19
1.
Western Blot - 2_Electrophoresis [Video from GeneTex]
-
00:12
2.
Clean the glass plates
-
00:22
3.
Assemble the gel apparatus
-
00:08
4.
Leaking test
-
00:08
4.1
Pour off H2O
-
00:12
5.
Preparation of separating gel
-
00:08
5.1
Add H2O
-
00:10
5.2
Add 1.5M Tris (pH8.8)
-
00:11
5.3
Add 30% polyacrylamide
-
00:13
5.4
Add 10% SDS
-
00:13
5.5
Add 10% ammonium persulfate (APS)
-
00:17
5.6
Add TEMED
-
00:11
5.7
Add the mixture into the gap between the glass plates
-
00:05
6.
Overlay with 75% EtOH
-
00:00
6.1
Remove 75% EtOH
-
00:03
6.2
Remove 75% EtOH
-
00:04
7.
Waiting for 30-60 min
-
00:00
8.
Preparation of stacking gel
-
00:20
8.1
Add H2O
-
00:07
8.2
Add 1M Tris (pH6.8)
-
00:11
8.3
Add 30% polyacrylamide
-
00:10
8.4
Add 10% SDS
-
00:11
8.5
Add 10% ammonium persulfate (APS)
-
00:15
8.6
Add TEMED
-
00:08
8.7
Add the mixture into the gap between the glass plates
-
00:16
9.
Insert a clean comb into the stacking gel solution
-
00:04
10.
Waiting for 15-30 min
-
00:17
11.
Remove the comb gently
-
00:00
12.
Running the gel
-
00:20
12.1
Assemble the electrophoresis apparatus
-
00:07
12.2
Add running buffer
-
00:10
12.3
Vortex samples & protein marker
-
00:03
12.4
Centrifuge samples & protein marker
-
00:15
12.5
Loading samples & protein marker
-
00:05
12.6
Attach the electrophoresis apparatus to the power supply
-
00:06
12.7
Apply the voltage of 80V to the gel, run 15 min
-
00:16
12.8
After the dye front has moved into the stacking gel, increase the voltage to 140V and run 50 min
- Index
- Notes
- Comment
- Fullscreen
Western Blot - 2_Electrophoresis [Video from GeneTex]
Duration: 06:35, Browse: 782, Last Updated: 2018-12-19
Play Video: http://llai.cm.ntu.edu.tw/media/170
Keypoint
- 1.
- 2.Leaking testConfirmed that there is no residual H2O between glass plates
- 3.Preparation of seperating gel<15 kDa 20%
15-50 kDa 15%
>50 kDa 12%
- 4.Preparation of seperating gel.
- 5.Preparation of stacking gel.
- 6.Insert a clean comb into the stacking gel solution.
- 7.Add running buffer.
-
00:19
1.
Western Blot - 2_Electrophoresis [Video from GeneTex]
-
00:12
2.
Clean the glass plates
-
00:22
3.
Assemble the gel apparatus
-
00:08
4.
Leaking test
-
00:08
4.1
Pour off H2O
-
00:12
5.
Preparation of separating gel
-
00:08
5.1
Add H2O
-
00:10
5.2
Add 1.5M Tris (pH8.8)
-
00:11
5.3
Add 30% polyacrylamide
-
00:13
5.4
Add 10% SDS
-
00:13
5.5
Add 10% ammonium persulfate (APS)
-
00:17
5.6
Add TEMED
-
00:11
5.7
Add the mixture into the gap between the glass plates
-
00:05
6.
Overlay with 75% EtOH
-
00:00
6.1
Remove 75% EtOH
-
00:03
6.2
Remove 75% EtOH
-
00:04
7.
Waiting for 30-60 min
-
00:00
8.
Preparation of stacking gel
-
00:20
8.1
Add H2O
-
00:07
8.2
Add 1M Tris (pH6.8)
-
00:11
8.3
Add 30% polyacrylamide
-
00:10
8.4
Add 10% SDS
-
00:11
8.5
Add 10% ammonium persulfate (APS)
-
00:15
8.6
Add TEMED
-
00:08
8.7
Add the mixture into the gap between the glass plates
-
00:16
9.
Insert a clean comb into the stacking gel solution
-
00:04
10.
Waiting for 15-30 min
-
00:17
11.
Remove the comb gently
-
00:00
12.
Running the gel
-
00:20
12.1
Assemble the electrophoresis apparatus
-
00:07
12.2
Add running buffer
-
00:10
12.3
Vortex samples & protein marker
-
00:03
12.4
Centrifuge samples & protein marker
-
00:15
12.5
Loading samples & protein marker
-
00:05
12.6
Attach the electrophoresis apparatus to the power supply
-
00:06
12.7
Apply the voltage of 80V to the gel, run 15 min
-
00:16
12.8
After the dye front has moved into the stacking gel, increase the voltage to 140V and run 50 min
- Location
-
- Folder name
- Western blot (Genetex版本)
- Author
- 賴亮全
- Branch
- 賴亮全教授
- Created
- 2018-12-19 18:19:09
- Last Updated
- 2018-12-19 23:32:42
- Browse
- 782
- Duration
- 06:35
- Reference
- 1