Team:Wash U/Calendar-Home/25 June 2009

From 2009.igem.org

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== Wednesday June 24, 2009 ==
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{{WashUback}}
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== Thursday June 25, 2009 ==
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-Minipreps of biobrick plasmid DNA (cultures from Th. 6/18)
-Minipreps of biobrick plasmid DNA (cultures from Th. 6/18)
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-Overnight PCR with miniprepped DNA
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-Overnight PCR with miniprepped DNA, to check for inserts
DNA nanospec concentrations (ng/microliter):
DNA nanospec concentrations (ng/microliter):
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PCR (1) labels
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'''PCR,      Miniprep,          Part #,            Conc(ng/microliter),      Plasmid,    Plate'''
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(A),            1,                    BBa_I15010,    22.2,      pSB2K3,  LBK50
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(B),              2,                  BBa_R0082,      51.4,      pSB1A2,  LBA50
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(C),            4,                    BBa_B0015,      75.9,      pSB1AK3, LBA50
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(D),            5,                    BBa_J04450,      81.4,      pSB1K3,    LBK50
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(E),              7,                  BBa_B0034,        49.6,      pSB1A2,  LBA50
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(F),              9,                  BBa_E0240,        62.9,      pSB1A2,  LBA50
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(G),            10,                BBa_J13002,        52.2,    pSB1A2,    LBA50
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'''PCR Reactions to check Biobricks in Plasmids'''
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AccuTaq LA
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[[1X]]                                          [[8X Master Mix]]
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5 uL of 10x Buffer                              40 uL
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2.5 uL of 10mM dNTP mix                        20 uL
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5 uL of template DNA (about 40 ng/uL)         
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1 uL DMSO                                      8 uL
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.5 uL 100X Primer F (Biobrick F)                4 uL
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.5 uL 100X Primer R (Biobrick R)                4 uL
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.5 uL Accutaq Enzyme                            4 uL
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35 uL H20                                      280 uL
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50 uL
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PCR labels:
 
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1(A)
 
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2(B)
 
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4(C)
 
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5(D)
 
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7(E)
 
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9(F)
 
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10(G)
 
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[[Team:Wash_U/Calendar-Home/23 June 2009|Previous entry]]</div>
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[[Team:Wash_U/Calendar-Home/24 June 2009|Previous entry]]</div>
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  [[Team:Wash_U/Calendar-Home/25 June 2009|Next entry]]</div>
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  [[Team:Wash_U/Calendar-Home/26 June 2009|Next entry]]</div>
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Latest revision as of 16:46, 6 July 2009



Thursday June 25, 2009

-Restreaked sphaeroides plates and plated from liquid cultures. We are working on getting rid of contamination on the earlier plates and in cultures.

-Waiting on supplies from Sigma Aldrich!

-Working on our webpage.... We have viewers from 4 continents!

-Minipreps of biobrick plasmid DNA (cultures from Th. 6/18)

-Overnight PCR with miniprepped DNA, to check for inserts

DNA nanospec concentrations (ng/microliter):

1) 22.2

2) 51.4

3) 135

4) 75.9

5) 81.4

6) 31.1

7) 49.6

8) 35.9

9) 62.9

10) 52.2

11) 134

12) 45.5


PCR (1) labels

PCR, Miniprep, Part #, Conc(ng/microliter), Plasmid, Plate

(A), 1, BBa_I15010, 22.2, pSB2K3, LBK50

(B), 2, BBa_R0082, 51.4, pSB1A2, LBA50

(C), 4, BBa_B0015, 75.9, pSB1AK3, LBA50

(D), 5, BBa_J04450, 81.4, pSB1K3, LBK50

(E), 7, BBa_B0034, 49.6, pSB1A2, LBA50

(F), 9, BBa_E0240, 62.9, pSB1A2, LBA50

(G), 10, BBa_J13002, 52.2, pSB1A2, LBA50


PCR Reactions to check Biobricks in Plasmids AccuTaq LA

1X 8X Master Mix

5 uL of 10x Buffer 40 uL 2.5 uL of 10mM dNTP mix 20 uL 5 uL of template DNA (about 40 ng/uL) 1 uL DMSO 8 uL .5 uL 100X Primer F (Biobrick F) 4 uL .5 uL 100X Primer R (Biobrick R) 4 uL .5 uL Accutaq Enzyme 4 uL 35 uL H20 280 uL

50 uL


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