August/8 August 2009

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Today we carried out the following procedures (in rough chronological order):

1. Checked the cell cultures transformed and plated out yesterday (8/7) for colony formation and made an approximate count of the number of colonies.

(plate number)-(location on plate) (no. of colonies)

            [http://partsregistry.org/wiki/index.php?title=Part:BBa_K081020 2-12H]                        none
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0071 1-12C]                        none
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0079 1-12A]                        none
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_F1610 2-24G]                        none
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118013 2-18F]                        none
                                               
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0062 1-6O]                         ~10
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0077 1-10K]                        ~10
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_I14017 1-18L]                        ~10

2. Conducted a 'miniprep' to harvest and check concentration of plasmids from 10 sample. (incubated since yesterday 8/7~) Result of Nanodrop concentration check:

(sample No.) (concentration) [260/280] [260/230]

    [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015 B0015](1-23L)           24.9 ng/uL     1.77      1.77
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118002 K118002](2-16F)       36.0 ng/uL     1.77      1.99
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_I0462 I0462](1-8O)            19.3 ng/uL     1.65      1.59
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 B0034](1-2M)            18.4 ng/uL     1.64      2.01
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_I742158 I742158](1-19B)       40.9 ng/uL     1.56      0.83
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118003 K118003](2-16H)       27.7 ng/uL     1.68      1.38
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0076 C0076](1-14D)            9.3 ng/uL     1.66      2.31
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0179 C0179](2-8M)             9.9 ng/uL     1.65      1.90
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_S03878 S03878](2-16M)         24.6 ng/uL     1.72      1.24
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0077 C0077](1-14F)            2.5 ng/uL     1.11      0.25   failure?
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_I1466 I1466](1-23J)            failure

3. gel electrophoresis

	uL
EcoRI	1
SpeI	1
10*Buffer No.2	2
DNA (EpsE)	5
total	20

↓
37°C,22hr
↓
failure　(SYBR Safe : 30min???)