Team:University of Ottawa/25 June 2008

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__TOC__
__TOC__
==Today in the Lab==
==Today in the Lab==
Line 14: Line 82:
::<li> Cleaned out all DNA that will not be used from our box
::<li> Cleaned out all DNA that will not be used from our box
::<li> Our names are getting complicated so I am renaming our DNA using the following legend (all other DNA was discarded)
::<li> Our names are getting complicated so I am renaming our DNA using the following legend (all other DNA was discarded)
-
:::597        > 7B   > 7
+
:::{| class="wikitable"
-
:::598         > 8B   > 8
+
|-
-
:::S1           > SB   > S
+
| '''Plasmid''' || '''Strain''' || '''New Name'''
-
:::D12       > DA   > D
+
|-
-
:::T123     > TB   > T
+
| 587 || 7B || 7
-
:::pSSA42 > 42B > 42  
+
|-
 +
| 598 || 8B || 8
 +
|-
 +
| 600 || 0A || 0
 +
|-
 +
| 601 || 1C || 1
 +
|-
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| S1 || SB || S
 +
|-
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| D12 || DA || D
 +
|-
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| T123 || TB || T
 +
|-
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| pSSA42 || 42B || 42
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|}
:'''PCR amplification of S, D, T, 7 and 8'''
:'''PCR amplification of S, D, T, 7 and 8'''
-
::<li> S, D, and T plasmids were PCR amplified using primers F69 and F70
+
::<li> S, D, and T plasmids were PCR amplified overnight using primers F69 and F70
 +
::<li> PCR program for 0&1 was: PHUSION
 +
::<li> PCR program for S&D&T was: PHN1
 +
::<li> The 0 and 1 plasmids were amplified according to the following chart: (Primer F66 is :Int TetR Repl F (*PstI) (B). primer F67 is Int GFP Repl F(*SphI) (A), and primer F68 is:Int SSRE R(*ClaI )
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:::{| class="wikitable"
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|-
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| '''Sample''' || '''F66''' || '''F67''' || '''F68'''
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|-
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| 0A ||    || Y || Y
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|-
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| 1A ||    || Y || Y
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|-
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| 0B || Y ||    || Y
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|-
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| 1B || Y ||    || Y
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Latest revision as of 23:38, 28 October 2008

Untitled Document

 

 

Contents

Today in the Lab

Matt

PCR Amplification
  • F60, F61 primers used for PTP2 cassette. I also labelled all the primers we received that we will be using for the next few months.
  • I decided to use the Kaern Lab protocol for PCR Amplification.
  • The PTP2 PCR product was then run on a gel - gel results were as expected with 2275 size band.
    • Glycerol Stock
  • Glycerol stock of plasmids D12, S1, T123, pSS042 with LB + glycerol was performed.

    • Dan

    Ran gel of pSSA42, S1, D12, T123 plasmids
  • This time the marker was added and expected results were obtained. SB, DA, TB, and 42B colonies were chosen for glycerol stock.
    • Cleaned out DNA box
  • Cleaned out all DNA that will not be used from our box
  • Our names are getting complicated so I am renaming our DNA using the following legend (all other DNA was discarded)
    • Plasmid Strain New Name
    587 7B 7
    598 8B 8
    600 0A 0
    601 1C 1
    S1 SB S
    D12 DA D
    T123 TB T
    pSSA42 42B 42
    PCR amplification of S, D, T, 7 and 8
  • S, D, and T plasmids were PCR amplified overnight using primers F69 and F70
  • PCR program for 0&1 was: PHUSION
  • PCR program for S&D&T was: PHN1
  • The 0 and 1 plasmids were amplified according to the following chart: (Primer F66 is :Int TetR Repl F (*PstI) (B). primer F67 is Int GFP Repl F(*SphI) (A), and primer F68 is:Int SSRE R(*ClaI )
    • Sample F66 F67 F68
    0A Y Y
    1A Y Y
    0B Y Y
    1B Y Y
    Retrieved from "http://2008.igem.org/Team:University_of_Ottawa/25_June_2008"