Team:UNIPV-Pavia/Calendar/August/settimana1

From 2010.igem.org

(Difference between revisions)
(August, 3rd)
(August, 3rd)
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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.
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.
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{| align="center"
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|[[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]]
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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.
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.
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Imgs
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{|align="center"
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|[[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>]]
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|[[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>]]
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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):
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):

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
MG1655
<partinfo>BBa_J72008</partinfo> MC1061
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

Calendar

August

week 1

week 2

week 3

week 4

week 5