Team:UNIPV-Pavia/Calendar/August/settimana1

AUGUST: WEEK 1

August, 2nd

Miniprep and quantification at Nanodrop of:

• I20-1: 98,2 ng/ul
• I20-2: 63,6 ng/ul
• I20-3: 41,5 ng/ul
• I21-1: 45 ng/ul
• I21-2: 45 ng/ul
• I21-3: 54 ng/ul

These samples were prepared and sent (400ng) to BMR Genomics for sequencing.

The following parts were resuspended from iGEM 2010 Distribution Kit:

• <partinfo>BBa_R0062</partinfo> (Plate 1, Well 6O)
• <partinfo>BBa_K081009</partinfo> (Plate 2, Well 10N)

both in vector <partinfo>pSB1A2</partinfo>.

Transformation (1ul) of the following parts (resuspended/already miniprepped):

Transformed cells were plated on proper LB+Amp agar plates and grown ON at right temperature:

August, 3rd

Check for plates grown ON: all plates showed colonies.

A single colony was picked from <partinfo>BBa_R0062</partinfo> and <partinfo>BBa_K081009</partinfo> plates and inoculated in 1 ml LB+Amp 100 ug/ml and incubated 37°C, 220 rmp for glycerol stock.

<partinfo>BBa_R0062</partinfo> plate

<partinfo>BBa_K081009</partinfo> plate

Since plates left showed small colonies they were let grow until late afternoon; than single colonies were picked and inoculated into 5 ml LB+Amp 50 ug/ml and incubated ON at 37°C, 220 rpm.

MC1061 transformed with pAH123	MC1061 transformed with <partinfo>BBa_J72008</partinfo>
MG1655 transformed with pAH123	MG1655 transformed with <partinfo>BBa_J72008</partinfo>

PCR from the following colonies (this is a test to check the efficiency of primers and it will be our negative control for E. coli integration screenings):

• MC1061-1
• MC1061-2
• MG1655-1
• MG1655-2
• Blank (Nothing)

using our primers synthesized to check attPhi80 E. coli genomic integration.

Gel run of amplified DNA showed for every sample the expected distance between primers of a strain with nothing integrated in attPhi80 site (~XXX bp), but unfortunately we forgot to take a picture of the gel ;(

August, 4th

August, 5th

August, 6th

August, 7th

August, 8th