Team:UNIPV-Pavia/Calendar/July/settimana4

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(Difference between revisions)
(→July, 19th)
(→July, 20th)
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Single colonies were picked from plates and grown in LB+Cm at proper concentration. MG1655 colonies were let grow in LB+Cm to check if they integrated the Cm resistance of RING.
Single colonies were picked from plates and grown in LB+Cm at proper concentration. MG1655 colonies were let grow in LB+Cm to check if they integrated the Cm resistance of RING.
 +
 +
MiniPrep was performed on cultures incubated yesterday, with following yields:
 +
 +
{| border='1' align='center'
 +
|''Culture''||''Quantification''
 +
|-
 +
|I10-1 || 117.8 ng/ul
 +
|-
 +
| I12-2||105,9 ng/ul
 +
|-
 +
| I3-1 || 166,0 ng/ul
 +
|-
 +
|I14-1 || 116,8 ng/ul
 +
|-
 +
| I17-1 || 189,5 ng/ul
 +
|-
 +
|<partifno>BBa_J23110</partinfo>||169,9 ng/ul
 +
|}
 +
 +
We retrieved from our freezer the following MiniPreps, with quantifications:
 +
 +
 +
{| border='1' align='center'
 +
|''Culture''||''Quantification''
 +
|-
 +
| I4C5 || 276 ng/ul
 +
|-
 +
| I16-1||68,4 ng/ul
 +
|-
 +
| I18-1 || 63,6 ng/ul
 +
|-
 +
| I19-1 || 58,8 ng/ul
 +
|}
 +
 +
Digestion of:
 +
 +
{| border="1" align='center'
 +
|  ''Culture''    ||  ''Kind''  ||  ''Final reaction volume (ul) '' ||  ''DNA (ul)''  ||  ''H20 (ul)''  ||  ''Enzyme 1''  ||  ''Enzyme 2'' ||  ''Buffer H''
 +
|-
 +
|    <partinfo>BBa_J23110</partinfo>  ||  Vector || 25 ||  6  ||  14,5  ||      1 SpeI  || 1 PstI  || 2,5
 +
|-
 +
|  I3-1  ||  Vector || 25 || 10,5  ||  10  ||      1 XbaI  || 1 PstI  || 2,5
 +
|-
 +
|  4C5  ||  Vector || 25 ||  3,6  ||  16,9  ||      1 EcoRI  || 1 PstI  || 2,5
 +
|-
 +
|  I14-1  ||  Insert || 25 ||  12,8  ||  7,7  ||      1 EcoRI  || 1 PstI  || 2,5
 +
|-
 +
|  I16-1  || Insert || 25 ||  12,5  ||  8  ||      1 EcoRI  || 1 PstI  || 2,5
 +
|-
 +
|  I17-1 ||  Insert || 25 ||  8  ||  12,5  ||      1 EcoRI  || 1 PstI  || 2,5
 +
|-
 +
|  I18-1  ||  Insert || 25 ||  13  ||  7,5  ||      1 EcoRI  || 1 PstI  || 2,5
 +
|-
 +
|  I19-1 ||  Insert || 25 ||  13  ||  7,5  ||      1 EcoRI  || 1 PstI  || 2,5
 +
|-
 +
|  I12-2  ||  Insert || 25 ||  14  ||  6,5  ||      1 EcoRI  || 1 PstI  || 2,5
 +
|-
 +
|  I10-1 ||  Insert || 25 ||  12,5  ||  8  ||      1 EcoRI  || 1 PstI  || 2,5
 +
|}
==July, 21st==
==July, 21st==

Revision as of 16:44, 29 July 2010
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JULY: WEEK 4


July, 19th

TECAN test showed that no RFP was produced from our parts, so all parts are potentially correct! For this reason we decided to sequence:

  • I14-1 (Forward)
  • I16-1 (Forward)
  • I17-1 (Forward)
  • I18-1 (Forward)
  • I19-1 (Forward)

We also sequenced:

  • I74C5-2 (Forward and Reverse)
  • I84C5-2 (Forward and Reverse)
  • I12-2 (Forward and Reverse)

These samples were prepared for sequencing (DNA was essicated) and sent to BMR genomics.

Trasformation of RING into:

  • BW25141 (pir+)
  • BW25142 (pir116)
  • BW23474 (pir116)
  • DH5alpha
  • MG1655

Cultures were plated on:

  • BW2514: Cm 34ug/ml
  • BW25142: Cm 34ug/ml
  • BW234741: Cm 34ug/ml
  • DH5alpha: Cm 12,5ug/ml
  • MG1655: Cm 12,5ug/ml


Inoculum of:

I10-1I12-2I17-1
I14-1<partinfo>BBa_J23110</partinfo>I3-1

in 5ml LB+Amp. Cultures were grown ON 37°C 220 rpm.

July, 20th

BW23474 transformed with <partinfo>BBa_J72007</partinfo>
DH5alpha transformed with <partinfo>BBa_J72007</partinfo>

Results for plates incubated ON, 37° C:

  • BW23474 (pir116): showed colonies
  • BW25141 (pir+): showed colonies
  • BW25142 (pir116): showed colonies
  • DH5alpha: didn't show colonies
  • MG1655: didn't show colonies (even if there were a very few colonies that we suppose integrated the resistance of RING to survive - or the plate antibiotic wasn't homogeneous)


BW23474 transformed with RING
BW25141 transformed with RING
BW25142 transformed with RING
DH5alpha transformed with RING
MG1655 transformed with RING

Single colonies were picked from plates and grown in LB+Cm at proper concentration. MG1655 colonies were let grow in LB+Cm to check if they integrated the Cm resistance of RING.

MiniPrep was performed on cultures incubated yesterday, with following yields:

CultureQuantification
I10-1 117.8 ng/ul
I12-2105,9 ng/ul
I3-1 166,0 ng/ul
I14-1 116,8 ng/ul
I17-1 189,5 ng/ul
<partifno>BBa_J23110</partinfo>169,9 ng/ul

We retrieved from our freezer the following MiniPreps, with quantifications:


CultureQuantification
I4C5 276 ng/ul
I16-168,4 ng/ul
I18-1 63,6 ng/ul
I19-1 58,8 ng/ul

Digestion of:

Culture Kind Final reaction volume (ul) DNA (ul) H20 (ul) Enzyme 1 Enzyme 2 Buffer H
<partinfo>BBa_J23110</partinfo> Vector 25 6 14,5 1 SpeI 1 PstI 2,5
I3-1 Vector 25 10,5 10 1 XbaI 1 PstI 2,5
4C5 Vector 25 3,6 16,9 1 EcoRI 1 PstI 2,5
I14-1 Insert 25 12,8 7,7 1 EcoRI 1 PstI 2,5
I16-1 Insert 25 12,5 8 1 EcoRI 1 PstI 2,5
I17-1 Insert 25 8 12,5 1 EcoRI 1 PstI 2,5
I18-1 Insert 25 13 7,5 1 EcoRI 1 PstI 2,5
I19-1 Insert 25 13 7,5 1 EcoRI 1 PstI 2,5
I12-2 Insert 25 14 6,5 1 EcoRI 1 PstI 2,5
I10-1 Insert 25 12,5 8 1 EcoRI 1 PstI 2,5

July, 21st

July, 22nd

July, 23rd

July, 24th

July, 25th

Calendar

July

week 1

week 2

week 3

week 4

week 5

Retrieved from "http://2010.igem.org/Team:UNIPV-Pavia/Calendar/July/settimana4"