Team:Rice University/Notebook/9 June 2008

From 2008.igem.org

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(Monday 9 June)
(Monday 9 June)
 
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****Tm = 81.7 C    TaOpt: 60.4 C    GC: 56.3
****Tm = 81.7 C    TaOpt: 60.4 C    GC: 56.3
****area of interest = 20833
****area of interest = 20833
-
****sense primer: GAGACGAATGCCAGGTCATCTGAAA <---------------primer name: ATTP L
+
****sense primer: GAGACGAATGCCAGGTCATCTGAAA <---------------primer name: STF L
****length: 25    Tm = 60.3 C    GC = 48.0
****length: 25    Tm = 60.3 C    GC = 48.0
-
****antisense primer: AAATCTGGATCATTCCCGAGCGCTG <-----------primer name: ATTP R
+
****antisense primer: AAATCTGGATCATTCCCGAGCGCTG <-----------primer name: STF R
****length: 25    Tm = 64.9 C    GC = 52.0
****length: 25    Tm = 64.9 C    GC = 52.0
***Primer Set 2:
***Primer Set 2:
Line 16: Line 16:
****Tm = 70.7 C  TaOpt: 51.4 C    GC: 31.7
****Tm = 70.7 C  TaOpt: 51.4 C    GC: 31.7
****area of interest = 23539
****area of interest = 23539
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****sense primer: ATCACATCGTCACCCATTGGATTGT <---------------primer name: STF L
+
****sense primer: ATCACATCGTCACCCATTGGATTGT <---------------primer name: ATTP L
****length: 25    Tm = 60.1 C    GC = 44.0
****length: 25    Tm = 60.1 C    GC = 44.0
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****antisense primer: CGATTTAGAAATGTATAGCGAGGCA <-----------primer name: STF R
+
****antisense primer: CGATTTAGAAATGTATAGCGAGGCA <-----------primer name: ATTP R
****length: 25    Tm = 55.9 C    GC = 40.0
****length: 25    Tm = 55.9 C    GC = 40.0
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*Taylor Stevenson
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**XL1-Blue MR cells were prepared for phage infection as specified in packaging manual [https://static.igem.org/mediawiki/2008/6/6a/Packaging_Extract.pdf] and infected with phage at roughly a 1/10 pfu/cfu ratio.  Resulting phage/bacteria mixture was incubated @ 37*C for 20 min and then streaked onto an LB plate.  Plate was incubated @ 30*C O/N. 
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**'''Result'''-approximately 100 possibly lysogenic colonies grew on plate.
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<BR><BR><BR><BR>
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{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
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!align="center"|[[Team:Rice_University|Home]]
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!align="center"|[[Team:Rice_University/Team|The Team]]
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!align="center"|[[Team:Rice_University/Project|The Project]]
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!align="center"|[[Team:Rice_University/Parts|Parts Submitted to the Registry]]
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!align="center"|[[Team:Rice_University/Modeling|Modeling]]
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!align="center"|[[Team:Rice_University/Notebook|Notebook]]
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|}

Latest revision as of 20:42, 16 July 2008

Monday 9 June

  • Selim Sheikh:
    • Designed 2 sets of sequencing primers (using Vector NTI Advance 10 http://www.invitrogen.com/site/us/en/home/LINNEA-Online-Guides/LINNEA-Communities/Vector-NTI-Community/Sequence-analysis-and-data-management-software-for-PCs.html) to be used in PCR of lambda DNA to amplify target regions:
      • Primer Set 1:
        • product of length 538
        • contains region of the molecule from 20481 to 21018
        • Tm = 81.7 C TaOpt: 60.4 C GC: 56.3
        • area of interest = 20833
        • sense primer: GAGACGAATGCCAGGTCATCTGAAA <---------------primer name: STF L
        • length: 25 Tm = 60.3 C GC = 48.0
        • antisense primer: AAATCTGGATCATTCCCGAGCGCTG <-----------primer name: STF R
        • length: 25 Tm = 64.9 C GC = 52.0
      • Primer Set 2:
        • product of length 309
        • contains region of the molecule from 23341 to 23649
        • Tm = 70.7 C TaOpt: 51.4 C GC: 31.7
        • area of interest = 23539
        • sense primer: ATCACATCGTCACCCATTGGATTGT <---------------primer name: ATTP L
        • length: 25 Tm = 60.1 C GC = 44.0
        • antisense primer: CGATTTAGAAATGTATAGCGAGGCA <-----------primer name: ATTP R
        • length: 25 Tm = 55.9 C GC = 40.0
  • Taylor Stevenson
    • XL1-Blue MR cells were prepared for phage infection as specified in packaging manual [1] and infected with phage at roughly a 1/10 pfu/cfu ratio. Resulting phage/bacteria mixture was incubated @ 37*C for 20 min and then streaked onto an LB plate. Plate was incubated @ 30*C O/N.
    • Result-approximately 100 possibly lysogenic colonies grew on plate.





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