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

(→‎Analyzing SDS PAGE Gel results)
(→‎Analyzing SDS PAGE Gel results)
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*Danny ran the SDS PAGE gel for the honeybee purification protocol on 7/30 and Megan stained the gel for visualization of 7/31.
 
*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.
 
**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 [[File:UCLA Bl21honeybee.jpg|none|thumb|900px|'''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, BSA positive control, and ladder (see table)]]
+
*Here is the gel [[File:UCLA Bl21honeybee.jpg|none|thumb|900px|'''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 [https://2015.igem.org/Team:UCLA/Notebook/Honeybee_Silk/29_July_2015#Honeybee_Inclusion_Body_Purification 7/29] for more detailed description of when each fraction was taken.
 
*Refer to [https://2015.igem.org/Team:UCLA/Notebook/Honeybee_Silk/29_July_2015#Honeybee_Inclusion_Body_Purification 7/29] for more detailed description of when each fraction was taken.
 +
*{| class="wikitable" style="text-align:center; width:400px; height:200px;"
 +
|+
 +
|-
 +
! F1
 +
! Full cell lysate using BugBuster lysis system
 +
|-
 +
! S1
 +
| Supernatant after spinning down full cell lysate
 +
|-
 +
! F2
 +
| Full cell lysate #2 after resuspending pellet in Bugbuster
 +
|-
 +
! F3
 +
| Full cell lysate #3 after adding lysozyme and DNase
 +
|-
 +
! S2
 +
| Supernatant after first wash step.
 +
|-
 +
 +
!S3
 +
|Supernatant after second wash step
 +
|-
 +
! Final product
 +
| Purified inclusion body in SDS
 +
|-
 +
! positive control
 +
| BSA positive control
 +
|-
 +
 +
!ladder
 +
|BioRad dual color ladder
 +
|}

Revision as of 18:37, 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.
  • {| class="wikitable" style="text-align:center; width:400px; height:200px;"

|+ |- ! F1 ! Full cell lysate using BugBuster lysis system |- ! S1 | Supernatant after spinning down full cell lysate |- ! F2 | Full cell lysate #2 after resuspending pellet in Bugbuster |- ! F3 | Full cell lysate #3 after adding lysozyme and DNase |- ! S2 | Supernatant after first wash step. |-

!S3 |Supernatant after second wash step |- ! Final product | Purified inclusion body in SDS |- ! positive control | BSA positive control |-

!ladder |BioRad dual color ladder |}