NYMU Taipei/Lab Notes/2007 9 27

From 2007.igem.org

(Difference between revisions)
Line 17: Line 17:
*** PCR product length without any insert = (3,189 - 3,049) + 176 = 316
*** PCR product length without any insert = (3,189 - 3,049) + 176 = 316
[[Image:NYMU Taipei 20070927 CinR-HSL-D-term plate culture test.jpg]]
[[Image:NYMU Taipei 20070927 CinR-HSL-D-term plate culture test.jpg]]
-
----
 
* '''References'''
* '''References'''
** [http://partsregistry.org/Image:PsB1AK3_map.jpg vector map of pSB1AK3 and VF2 VR primers]
** [http://partsregistry.org/Image:PsB1AK3_map.jpg vector map of pSB1AK3 and VF2 VR primers]
 +
----
 +
* '''pOmpC glucose response testing by construct (pOmpC + EYFP)'''
 +
** design: 5 tubes for 0%, 1%, 3%, 5%, 10% extra glucose cultured in LB medium
 +
** tube with 0% extra glucose is treated as negative control
 +
{|
 +
!glucose (%)
 +
!O.D. 600
 +
!white light
 +
!green light
 +
|-
 +
|0
 +
|2.487
 +
|
 +
|
 +
|-
 +
|1
 +
|2.091
 +
|
 +
|
 +
|-
 +
|3
 +
|1.861
 +
|
 +
|
 +
|-
 +
|5
 +
|1.637
 +
|
 +
|
 +
|-
 +
|10
 +
|1.025
 +
|
 +
|
 +
|-
 +
|}

Revision as of 09:25, 27 September 2007

  • CinR+HSL+D-term check
    • After re-culture in 20-block plate,randomly pick 6 blocks from CinR+HSL+D-term plate
    • PCR check using VF2, VR primers
    • PCR total 50 uL, 10 uL for GEL separation
    • sample at each lane
      • 1Kb ladder
      • block #1 (insert band appears)
      • block #2 (insert band appears, and one extra band around 400 bp)
      • block #3 (insert band appears)
      • block #4 (insert band appears)
      • block #5 (insert band does not appear, only band around 400 bp)
      • block #6 (insert band does not appear, only band around 400 bp)
    • expect results: one band at 1,623 bp is our insert
      • insert = RBS+CinR (2,843 - 2,053) + RBS+HSL (2,783 - 2,053) + D-term (3,266 - 3,163)
      • insert = 790 + 730 + 103 = 1,623
    • extra band at 400 bp might be plasmid failed to be correctly ligated
      • PCR product length without any insert = (3,189 - 3,049) + 176 = 316

NYMU Taipei 20070927 CinR-HSL-D-term plate culture test.jpg

  • References
    • [http://partsregistry.org/Image:PsB1AK3_map.jpg vector map of pSB1AK3 and VF2 VR primers]
  • pOmpC glucose response testing by construct (pOmpC + EYFP)
    • design: 5 tubes for 0%, 1%, 3%, 5%, 10% extra glucose cultured in LB medium
    • tube with 0% extra glucose is treated as negative control
glucose (%) O.D. 600 white light green light
0 2.487
1 2.091
3 1.861
5 1.637
10 1.025
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