Team:UNIPV-Pavia/Calendar/July/settimana2

July, 4th

Digestions of previously purified plasmids were performed for ligations:

Plasmid	Kind	Purified Dna (μl)	H2O (μl)	Enzyme 1 (μl)	Enzyme 2 (μl)	Buffer H (μl)	Final Volume(μl)
<partinfo>BBa_C0060</partinfo>	Insert	16.5	4	1 EcoRI	1 SpeI	2.5	25
<partinfo>BBa_C0061</partinfo>	Insert	13.2	7.3	1 XbaI	1 PstI	2.5	25
<partinfo>BBa_K081022</partinfo>	Insert	15.7	4.8	1 EcoRI	1 PstI	2.5	25
<partinfo>BBa_B0030</partinfo>	Vector	13.2	7.3	1 SpeI	1 PstI	2.5	25
<partinfo>BBa_B0031</partinfo>	Vector	12.4	8.1	1 SpeI	1 PstI	2.5	25
<partinfo>BBa_B0032</partinfo>	Vector	9.5	11	1 SpeI	1 PstI	2.5	25
<partinfo>BBa_B0015</partinfo>	Vector	7.9	12.6	1 EcoRI	1 XbaI	2.5	25
<partinfo>pSB4C5</partinfo>	Vector	3	17.5	1 EcoRI	1 PstI	2.5	25
<partinfo>BBa_I13501</partinfo>	Insert	7.2	13.3	1 XbaI	1 PstI	2.5	25

Reaction samples were incubated at 37°C for three hours while a small-size and a medium-size agarose gel were prepared according to protocols.

In the afternoon gel electrophoresis was performed.

Small size gel

Medium size gel

As shown, in figure all clones were positive (apart from <partinfo>BBa_I13501</partinfo> run where we inexplicably didn't see any band), so we cut and purified the bands of interest.

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