Team:UNIPV-Pavia/Calendar/August/settimana1

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Revision as of 19:24, 3 August 2010

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_K081008</partinfo> (Plate 2, Well 10N)

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

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

Part	Strain
pAH123	MC1061
pAH123	MG1655
<partinfo>BBa_J72008</partinfo>	MC1061
<partinfo>BBa_J72008</partinfo>	MG1655
<partinfo>BBa_R0062</partinfo>	DH5-alpha
<partinfo>BBa_K081008</partinfo>	DH5-alpha

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

Part	Plate resistance	Temperature
pAH123	Amp 50 ug/ml	30°C
<partinfo>BBa_J72008</partinfo>
<partinfo>BBa_R0062</partinfo>	Amp 100 ug/ml	37°C
<partinfo>BBa_K081008</partinfo>

August, 3rd

Check for plates grown ON: all plates showed colonies.

A single colony was picked from <partinfo>BBa_R0062</partinfo> and <partinfo>BBa_K081008</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

File:UNIPV10 BBa K081008.jpg

<partinfo>BBa_K081008</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.

August, 4th

August, 5th

August, 6th

August, 7th

August, 8th

week 1

@@ Line 68: / Line 68: @@
 A single colony was picked from <partinfo>BBa_R0062</partinfo> and <partinfo>BBa_K081008</partinfo> plates and inoculated in 1 ml LB+Amp 100 ug/ml and incubated 37°C, 220 rmp for glycerol stock.
-Imgs
+{| align="center"
+|[[Image:UNIPV10_BBa_R0062.jpg|thumb|200px|center|<partinfo>BBa_R0062</partinfo> plate]] || [[Image:UNIPV10_BBa_K081008.jpg|thumb|200px|center|<partinfo>BBa_K081008</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.
-Imgs
+{|align="center"
+|-
+|[[Image:UNIPV10_MC1061_pah123.jpg|thumb|200px|center|MC1061 transformed with pAH123]] || [[Image:UNIPV10_MC1061_BBa_J72008.jpg|thumb|200px|center|MC1061 transformed with <partinfo>BBa_J72008</partinfo>]]
+|-
+|[[Image:UNIPV10_MG1655_pah123.jpg|thumb|200px|center|MG1655 transformed with pAH123]] || [[Image:UNIPV10_MG1655_BBa_J72008.jpg|thumb|200px|center|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):