Difference between revisions of "Team:UCLA/Notebook/Honeybee Silk/3 August 2015"

(Analyzing SDS PAGE Gel results)
(Analyzing SDS PAGE Gel results)
Line 294: Line 294:
 
|+  
 
|+  
 
|-
 
|-
! F1
+
! F1 (lane 1)
 
! Full cell lysate using BugBuster lysis system
 
! Full cell lysate using BugBuster lysis system
 
|-
 
|-
! S1
+
! S1 (lane 2)
 
| Supernatant after spinning down full cell lysate
 
| Supernatant after spinning down full cell lysate
 
|-
 
|-
! F2
+
! F2 (lane 3)
 
| Full cell lysate #2 after resuspending pellet in Bugbuster
 
| Full cell lysate #2 after resuspending pellet in Bugbuster
 
|-
 
|-
! F3
+
! F3 (lane 4)
 
| Full cell lysate #3 after adding lysozyme and DNase
 
| Full cell lysate #3 after adding lysozyme and DNase
 
|-
 
|-
! S2
+
! S2 (lane 5)
 
| Supernatant after first wash step.
 
| Supernatant after first wash step.
 
|-
 
|-
  
!S3
+
!S3 (lane 6)
 
|Supernatant after second wash step
 
|Supernatant after second wash step
 
|-
 
|-
! Final product
+
! Final product (lane 7)
 
| Purified inclusion body in SDS
 
| Purified inclusion body in SDS
 
|-
 
|-
! positive control
+
! positive control (lane 8)
 
| BSA positive control
 
| BSA positive control
 
|-
 
|-
  
!ladder
+
!ladder (lane 9)
 
|BioRad dual color ladder
 
|BioRad dual color ladder
 
|}
 
|}

Revision as of 18:42, 3 August 2015

Analyzing SDS PAGE Gel results

  • Danny ran the SDS PAGE gel for the honeybee purification protocol on 7/30 and Megan stained the gel for visualization of 7/31.
    • The gel was run, and then stored overnight in destain buffer due to logistical reasons before being stained the next day.
  • Here is the gel
    Fig. 1 Expected size of product is 40 kDa (lane 7). Lane 8 is BSA protein positive control, and lane 9 is Biorad dual color ladder. From lane 1-9 F1,S1,F2,F3,S2,S3, Final product, positive control, and ladder (see table)
  • Refer to 7/29 for more detailed description of when each fraction was taken.
F1 (lane 1) Full cell lysate using BugBuster lysis system
S1 (lane 2) Supernatant after spinning down full cell lysate
F2 (lane 3) Full cell lysate #2 after resuspending pellet in Bugbuster
F3 (lane 4) Full cell lysate #3 after adding lysozyme and DNase
S2 (lane 5) Supernatant after first wash step.
S3 (lane 6) Supernatant after second wash step
Final product (lane 7) Purified inclusion body in SDS
positive control (lane 8) BSA positive control
ladder (lane 9) BioRad dual color ladder