Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2014/11/06: Difference between revisions

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(Autocreate 2014/11/06 Entry for Haynes_Lab:Notebook/Characterizing_AHL_quorum_sensing_homologs)
 
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==mm/dd/yyyy==
==mm/dd/yyyy==
* Insert content here...
* Retried PCR of golden gate parts using Rene's primers, this time doing a Tm screen
 
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Reagent'''
| align="center" style="background:#f0f0f0;"|'''uL'''
|-
| ddH2O||33
|-
| HF buffer||10
|-
| primer F (10uM)||2.5
|-
| primer R (10uM)||2.5
|-
| dNTP (10mM)||1
|-
| Template K084007||0.5
|-
| Phusion polymerase ||0.5
|-
| Total ||50
|-
|}
<br>
<br>
 
*annealed for 30 seconds
*used Tm: 30, 32, 34, 36, 38
 
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Number'''
| align="center" style="background:#f0f0f0;"|'''Template'''
| align="center" style="background:#f0f0f0;"|'''F Primer'''
| align="center" style="background:#f0f0f0;"|'''R Primer'''
| align="center" style="background:#f0f0f0;"|'''Annealing T'''
| align="center" style="background:#f0f0f0;"|'''Expected length'''
|-
| 1||K084007 3/10/13 4-2-1||P069||P070||32||645
|-
| 2||K084007 3/10/13 4-2-2||P069||P070||32||645
 
|}
<br>
<br>
 
* Still no clear bands observed for any of the annealing temperatures.




Revision as of 14:03, 2 February 2015

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mm/dd/yyyy

  • Retried PCR of golden gate parts using Rene's primers, this time doing a Tm screen
Reagent uL
ddH2O 33
HF buffer 10
primer F (10uM) 2.5
primer R (10uM) 2.5
dNTP (10mM) 1
Template K084007 0.5
Phusion polymerase 0.5
Total 50



  • annealed for 30 seconds
  • used Tm: 30, 32, 34, 36, 38
Number Template F Primer R Primer Annealing T Expected length
1 K084007 3/10/13 4-2-1 P069 P070 32 645
2 K084007 3/10/13 4-2-2 P069 P070 32 645



  • Still no clear bands observed for any of the annealing temperatures.



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