Team:UNIPV-Pavia/Calendar/September

71° Bio-Lab - PCR from colony for I47 ligation.Tecan Test was performed on prepared samples.

72° Bio-Lab - Again PCR from colony for I47 ligation: I47-10..26.Tecan Test was performed on prepared samples.

73° Bio-Lab - Today we got our yeast strains.

74° Bio-Lab -

75° Bio-Lab -

76° Bio-Lab - Inoculum of I47, I48, I49 into 5 ml LB+Amp for TECAN experiment.

Inoculum of I44-2 and I51-3 into 5 ml LB+Cm34 to evaluate MiniPrep yields.

Inoculum of I44-2 and I51-3 into 5 ml LB+Cm12,5 to evaluate MiniPrep yields and for sequencing.

Inoculum for MiniPrep and sequencing of: <partinfo>BBa_J23100</partinfo>_4C5, <partinfo>BBa_J23101</partinfo>_4C5, <partinfo>BBa_J23105</partinfo>_4C5, <partinfo>BBa_J23106</partinfo>_4C5, <partinfo>BBa_J23110</partinfo>_4C5, <partinfo>BBa_J23114</partinfo>_4C5, <partinfo>BBa_J23116</partinfo>_4C5, <partinfo>BBa_J23118</partinfo>_4C5, I7-A and I8-5-D.

Inoculum of cultures for TECAN test on self-inducible promoters.

Glycerol stocks of MG43, MC42, MC43. Static TECAN test (absorbance, green fluorescence and red fluorescence) of these cultures.

Liquid PCR was performed on MG43, MC42, MC43 and MC1061. MiniPreps of I44-1:5 and I51-1:5 were performed.

77° Bio-Lab -MiniPreps of <partinfo>BBa_J23100</partinfo>_4C5, <partinfo>BBa_J23101</partinfo>_4C5, <partinfo>BBa_J23105</partinfo>_4C5, <partinfo>BBa_J23106</partinfo>_4C5, <partinfo>BBa_J23110</partinfo>_4C5, <partinfo>BBa_J23114</partinfo>_4C5, <partinfo>BBa_J23116</partinfo>_4C5, <partinfo>BBa_J23118</partinfo>_4C5, I7-A, I8-5-D, I44-2, I51-3, I44-2, I51-3 and I47 were performed. Trasformation of I42 and I43. Integration protocol's first day were performed.
13° Meeting - Updating about Bio-Lab activity.

78° Bio-Lab - Inoculum of: I31, I32, I35, I36, I37, I38, I40, I41 and I20 for second step of ligation of phasins/intein.

79° Bio-Lab - Minipresp of I31, I32, I35, I36, I37, I38, I40, I41 and I20 were performed.

80° Bio-Lab - Screening digestion of I44-2 and I51-3.

Gel run/cut for samples of previous day (I31, I32, I35, I36, I37, I38, I40, I41 and I20). Digestions were ligated in order to create: I53, I54, I55, I56, I57, I58.

81° Bio-Lab - MC42 and MC43 plates were transfered.

I42 and I43 have been transformed in MG008 strain, plated and incubated.

82° Bio-Lab - Trasformation of ligations I52, I54, I55, I56, I57 and I58. into E. coli DH5-alpha.

Inoculum of I26, I31 and I34.

Inoculum from single colony of MG42 and MC43.

83° Bio-Lab - Agar plates of I52, I54, I55, I56, I57 and I58 were checked.

Miniprep and quanfification of I26, I31 and I34. After 3 hour digestion, gel run/cut of samples was done. Gel exctraction and ligation was performed in order to obtain I59, I60, I61 and I53.

Inoculum of I47, I48, I49 into 5 ml LB+Amp for TECAN test of the following day.

Screening PCR on 3 colonies picked from MC42 and MC43 respectively.

Tecan Test was performed on prepared samples, after usual protocol.

84° Bio-Lab - Tecan Test was started to check GFP production by BioBricks in I47, I48, I49 cultures.

Transformation of I 53, I59, I60 and I61.

85° Bio-Lab - Screening through "colony PCR" for ligations I52, I53, I54, I55, I56, I57, I58, I59, I60 and I61 was performed.

Tecan Test was performed on prepared samples, after the usual protocol.

86° Bio-Lab - Miniprep and Nanodrop quantification for: I58a, I58b, I58c, I59a, I59b, I59c, I60a, I60b, I60c, I61a, I61b and I61c.

Digestion (for screening) and gel extraction for: I58a/b/c, I59a/b/c, I60a/b/c.

Digestion (only screening) for I61a/b/c.

87° Bio-Lab - Gel extraction and quantification of: I58, I59, I60. DNA was than stored at -20°C.

xx° Bio-Lab - I55 was transformed again into E. coli DH5-alpha and plated on LB+Amp agar plate, that was let grow ON at 37°C.

88° Bio-Lab -

89° Bio-Lab -

90° Bio-Lab

91° Bio-Lab -

92° Bio-Lab -

93° Bio-Lab -

xx° Bio-Lab -

94° Bio-Lab -
14° Meeting - Updating about Bio-Lab activity.

95° Bio-Lab -

96° Bio-Lab

97° Bio-Lab -