Glasgow/Wetlab/Week7

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== Week 7 ==
== Week 7 ==
=== Monday 13th August 2007 ===
=== Monday 13th August 2007 ===
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[[User:L.McLeay|Lynsey]] did colony PCR (see [[Glasgow/Wetlab/Protocols#Protocol 9:PCR| Protocol 9]]) on the following ''E. coli'' TOP10 transformants to check that their plasmids carry our expected inserts:
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<br>
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<br>
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{| align="center" border="1" cellpadding="5"
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|- align="center"
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|'''Plasmid''' || '''Expected Insert''' || '''Primers'''
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|- align="center"
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| pCR4 || Pu || [https://2007.igem.org/Glasgow/Wetlab/Orders Pu_Prefix_After] & [https://2007.igem.org/Glasgow/Wetlab/Orders Pu_Suffix_After]
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|- align="center"
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| pCR4 ||Pu || [https://2007.igem.org/Glasgow/Wetlab/Orders Pu_Prefix_Emma] & [https://2007.igem.org/Glasgow/Wetlab/Orders Pu_Suffix_Emma]
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|- align="center"
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| pCR4 ||Pr || [https://2007.igem.org/Glasgow/Wetlab/Orders Pr_Prefix_1] & [https://2007.igem.org/Glasgow/Wetlab/Orders Pr_Suffix_1]
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|- align="center"
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| pCR4 ||XylR || [https://2007.igem.org/Glasgow/Wetlab/Orders XylR_Prefix_1] & [https://2007.igem.org/Glasgow/Wetlab/Orders XylR_Suffix_1]
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|- align="center"
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| pCR4 ||XylR & Pr || [https://2007.igem.org/Glasgow/Wetlab/Orders Pr_Prefix_1] & [https://2007.igem.org/Glasgow/Wetlab/Orders XylR_Suffix_1]
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|}
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Overnight cultures were seeded from colonies that gave successful PCR results in order to miniprep the plasmids tomorrow.
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<li>
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[[User:MaijaP|Maija]] did colony PCR (see [[Glasgow/Wetlab/Protocols#Protocol 9:PCR| Protocol 9]]) on the following ''E. coli'' DB3.1 transformants to check that their plasmids carry our expected inserts:
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{| align="center" border="1" cellpadding="5"
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|- align="center"
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|'''Plasmid''' || '''Expected Insert''' || '''Primers'''
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|- align="center"
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| pSB3K5 (4/6B) || (*m*) B1 17 || [https://2007.igem.org/Glasgow/Wetlab/Orders (*m*)_for_1] & [https://2007.igem.org/Glasgow/Wetlab/Orders BBs_Methyl_2]
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|- align="center"
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| pSB1AC3 (3/20G) || (*m*) B1 17 || [https://2007.igem.org/Glasgow/Wetlab/Orders (*m*)_for_1] & [https://2007.igem.org/Glasgow/Wetlab/Orders BBs_Methyl_2]
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|- align="center"
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| pSB3K5 (4/6B) || (*m*) C5 24 || [https://2007.igem.org/Glasgow/Wetlab/Orders (*m*)_for_1] & [https://2007.igem.org/Glasgow/Wetlab/Orders (*m*)_rev_1]
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|- align="center"
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| pSB1AC3 (3/20G) || (*m*) || [https://2007.igem.org/Glasgow/Wetlab/Orders (*m*)_for_1] & [https://2007.igem.org/Glasgow/Wetlab/Orders (*m*)_rev_1]
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|- align="center"
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| pSB3K5 (4/6B) || DntR || [https://2007.igem.org/Glasgow/Wetlab/Orders DntR_Prefix_1] & [https://2007.igem.org/Glasgow/Wetlab/Orders DntR_Suffix_2]
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|- align="center"
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| pSB1AC3 (3/20G) || DntR || [https://2007.igem.org/Glasgow/Wetlab/Orders DntR_Prefix_1] & [https://2007.igem.org/Glasgow/Wetlab/Orders DntR_Suffix_2]
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|}
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Overnight cultures were seeded from colonies that gave successful PCR results in order to miniprep the plasmids tomorrow.
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<li>
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[[User:Mojs|Maia]] took the following BioBricks from the kit plates and transformed into ''E. coli'' TOP10 cells:
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{| align="center" border="1" cellpadding="5"
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|- align="center"
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|'''BioBrick Part''' || '''Function''' || '''Chassis''' || '''Plasmid''' || '''Kit Plate Location''' ''(Plate/Well)''
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|- align="center"
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|[http://partsregistry.org/Part:BBa_B0014 Bba_B0014] || Double terminator || ''E. coli''  TOP10 || pSB1AK3 || 1/1G
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|- align="center"
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|[http://partsregistry.org/Part:BBa_B0015 Bba_B0015] || Double terminator || ''E. coli''  TOP10 || pSB1AK3 || 1/1I
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|- align="center"
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| " || " || " || " || 3/3O
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|- align="center"
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|[http://partsregistry.org/Part:BBa_J61100 Bba_J61100] || RBS || ''E. coli''  TOP10 || pSB1A2 || 4/12N
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|- align="center"
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|[http://partsregistry.org/Part:BBa_J61101 Bba_J61101] || RBS || ''E. coli''  TOP10 || pSB1A2 || 4/12J
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|- align="center"
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|[http://partsregistry.org/Part:BBa_J61102 Bba_J61102] || RBS || ''E. coli''  TOP10 || pSB1A2 || 4/12L
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|- align="center"
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|[http://partsregistry.org/Part:BBa_J61103 Bba_J61103] || RBS || ''E. coli''  TOP10 || pSB1A2 || 4/12P
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|- align="center"
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|[http://partsregistry.org/Part:BBa_E0430 Bba_E0430] || EYFP + RBS + terminator || ''E. coli''  TOP10 || pSB1A2 || 1/11A
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|- align="center"
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|[http://partsregistry.org/Part:BBa_E0422 Bba_E0422] || ECFP + RBS + terminator + LVA tag || ''E. coli''  TOP10 || pSB1AK3 || 1/11G
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|- align="center"
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|[http://partsregistry.org/Part:BBa_E0432 Bba_E0432] || EYFP + RBS + terminator + LVA tag || ''E. coli''  TOP10 || pSB1A2 || 4/11C
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|}
=== Tuesday 14th August 2007 ===
=== Tuesday 14th August 2007 ===

Revision as of 20:11, 15 August 2007

Back To Main Page | Back To Wetlab Log

Contents

Week 7

Monday 13th August 2007

  1. Lynsey did colony PCR (see Protocol 9) on the following E. coli TOP10 transformants to check that their plasmids carry our expected inserts:

    Plasmid Expected Insert Primers
    pCR4 Pu Pu_Prefix_After & Pu_Suffix_After
    pCR4 Pu Pu_Prefix_Emma & Pu_Suffix_Emma
    pCR4 Pr Pr_Prefix_1 & Pr_Suffix_1
    pCR4 XylR XylR_Prefix_1 & XylR_Suffix_1
    pCR4 XylR & Pr Pr_Prefix_1 & XylR_Suffix_1

    Overnight cultures were seeded from colonies that gave successful PCR results in order to miniprep the plasmids tomorrow.

  2. Maija did colony PCR (see Protocol 9) on the following E. coli DB3.1 transformants to check that their plasmids carry our expected inserts:
    Plasmid Expected Insert Primers
    pSB3K5 (4/6B) (*m*) B1 17 (*m*)_for_1 & BBs_Methyl_2
    pSB1AC3 (3/20G) (*m*) B1 17 (*m*)_for_1 & BBs_Methyl_2
    pSB3K5 (4/6B) (*m*) C5 24 (*m*)_for_1 & (*m*)_rev_1
    pSB1AC3 (3/20G) (*m*) (*m*)_for_1 & (*m*)_rev_1
    pSB3K5 (4/6B) DntR DntR_Prefix_1 & DntR_Suffix_2
    pSB1AC3 (3/20G) DntR DntR_Prefix_1 & DntR_Suffix_2

    Overnight cultures were seeded from colonies that gave successful PCR results in order to miniprep the plasmids tomorrow.

  3. Maia took the following BioBricks from the kit plates and transformed into E. coli TOP10 cells:
    BioBrick Part Function Chassis Plasmid Kit Plate Location (Plate/Well)
    [http://partsregistry.org/Part:BBa_B0014 Bba_B0014] Double terminator E. coli TOP10 pSB1AK3 1/1G
    [http://partsregistry.org/Part:BBa_B0015 Bba_B0015] Double terminator E. coli TOP10 pSB1AK3 1/1I
    " " " " 3/3O
    [http://partsregistry.org/Part:BBa_J61100 Bba_J61100] RBS E. coli TOP10 pSB1A2 4/12N
    [http://partsregistry.org/Part:BBa_J61101 Bba_J61101] RBS E. coli TOP10 pSB1A2 4/12J
    [http://partsregistry.org/Part:BBa_J61102 Bba_J61102] RBS E. coli TOP10 pSB1A2 4/12L
    [http://partsregistry.org/Part:BBa_J61103 Bba_J61103] RBS E. coli TOP10 pSB1A2 4/12P
    [http://partsregistry.org/Part:BBa_E0430 Bba_E0430] EYFP + RBS + terminator E. coli TOP10 pSB1A2 1/11A
    [http://partsregistry.org/Part:BBa_E0422 Bba_E0422] ECFP + RBS + terminator + LVA tag E. coli TOP10 pSB1AK3 1/11G
    [http://partsregistry.org/Part:BBa_E0432 Bba_E0432] EYFP + RBS + terminator + LVA tag E. coli TOP10 pSB1A2 4/11C

    Tuesday 14th August 2007

    Wednesday 15th August 2007

    Thursday 16th August 2007

    === Friday 17th August 2007 ===
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