Team:Wash U/Calendar-Home/25 June 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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