Difference between revisions of "Repository:iGEM 2007 DNA distribution"

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We tested the antibiotic resistance of each part sent out in the Kit of Parts by growing it in broth containing each of our 4 antibiotics.  The test was done on a 384-deepwell plate filled with EZ Rich Defined Media which contained a cresol red indicator dye and the appropriate antibiotic.  Each quadrant of the test plate contains one of the four antibiotics into which the part is inoculated (see layout below).  After growing for 18hrs at 37C, images from each of the plates were collected.  Since the broth contains a cresol red indicator that changes as the pH of the media decreases, you can tell whether the part grew by noting which of the wells turned from red to yellow (as the cells grow, the media becomes more acidic).   
 
We tested the antibiotic resistance of each part sent out in the Kit of Parts by growing it in broth containing each of our 4 antibiotics.  The test was done on a 384-deepwell plate filled with EZ Rich Defined Media which contained a cresol red indicator dye and the appropriate antibiotic.  Each quadrant of the test plate contains one of the four antibiotics into which the part is inoculated (see layout below).  After growing for 18hrs at 37C, images from each of the plates were collected.  Since the broth contains a cresol red indicator that changes as the pH of the media decreases, you can tell whether the part grew by noting which of the wells turned from red to yellow (as the cells grow, the media becomes more acidic).   
  
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 +
'''384-deepwell test plate antibiotic quadrant layout:'''
  
 
{| align="left" border="1" cellpadding="4" style="text-align:center"
 
{| align="left" border="1" cellpadding="4" style="text-align:center"
−
|+ '''384-deepwell test plate antibiotic quadrant layout:'''
 
 
|-
 
|-
 
!width="50"| !! width="50"|1 !! width="50"|2 !! width="50"|3 !! width="50"|4 !! width="50"|...
 
!width="50"| !! width="50"|1 !! width="50"|2 !! width="50"|3 !! width="50"|4 !! width="50"|...
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[[Repository:iGEM 2007 DNA distribution/AB_Test/Source Plate 16|Source Plate 16]]
 
[[Repository:iGEM 2007 DNA distribution/AB_Test/Source Plate 16|Source Plate 16]]
 
|}
 
|}
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<br style="clear:both;"/>
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===Kit Plate Layout===
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 +
Each 384-well kit plate is a composition of 4 96-well source plates.  Below is a representation of which source plates went into which kit plate and in what order. 
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'''Kit Plate 1'''
 +
{| align="left" border="1" cellpadding="3" style="text-align:center"
 +
|-
 +
!width="50"| !! width="100"|1 !! width="100"|2 !! width="50"|...
 +
|-
 +
! A
 +
| style="background:#ffffff;" | Source Plate 1 || style="background:#ffffff;" | Source Plate 2 ||
 +
|-
 +
! B
 +
| style="background:#ffffff;" | Source Plate 3 || style="background:#ffffff;" | Source Plate 4 ||
 +
|-
 +
!...
 +
| || ||
 +
|}
 +
<br style="clear:both;"/>
 +
 +
 +
'''Kit Plate 2'''
 +
{| align="left" border="1" cellpadding="3" style="text-align:center"
 +
|-
 +
!width="50"| !! width="100"|1 !! width="100"|2 !! width="50"|...
 +
|-
 +
! A
 +
| style="background:#ffffff;" | Source Plate 5 || style="background:#ffffff;" | Source Plate 6 ||
 +
|-
 +
! B
 +
| style="background:#ffffff;" | Source Plate 7 || style="background:#ffffff;" | Source Plate 8 ||
 +
|-
 +
!...
 +
| || ||
 +
|}
 +
<br style="clear:both;"/>
 +
 +
 +
'''Kit Plate 3'''
 +
{| align="left" border="1" cellpadding="3" style="text-align:center"
 +
|-
 +
!width="50"| !! width="100"|1 !! width="100"|2 !! width="50"|...
 +
|-
 +
! A
 +
| style="background:#ffffff;" | 2006 Source Plate 1 || style="background:#ffffff;" | 2006 Source Plate 2 ||
 +
|-
 +
! B
 +
| style="background:#ffffff;" | 2006 Source Plate 3 || style="background:#ffffff;" | 2006 Source Plate 4 ||
 +
|-
 +
!...
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| || ||
 +
|}
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<br style="clear:both;"/>
 +
 +
 +
'''Kit Plate 4'''
 +
{| align="left" border="1" cellpadding="3" style="text-align:center"
 +
|-
 +
!width="50"| !! width="100"|1 !! width="100"|2 !! width="50"|...
 +
|-
 +
! A
 +
| style="background:#ffffff;" | Source Plate 5 (2007) || style="background:#ffffff;" | Source Plate 6 (2007) ||
 +
|-
 +
! B
 +
| style="background:#ffffff;" | Source Plate 7 (2007) || style="background:#ffffff;" | Source Plate 8 (2007) ||
 +
|-
 +
!...
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| || ||
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|}
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<br style="clear:both;"/>

Revision as of 18:03, 19 April 2007

From Mid-April to Early May, the staff at the Registry prepared the Kit of Parts that will be sent to the 2007 iGEM teams.

Here is some useful information about the steps taken in that process.

Quality Control

Antibiotic resistance:

We tested the antibiotic resistance of each part sent out in the Kit of Parts by growing it in broth containing each of our 4 antibiotics. The test was done on a 384-deepwell plate filled with EZ Rich Defined Media which contained a cresol red indicator dye and the appropriate antibiotic. Each quadrant of the test plate contains one of the four antibiotics into which the part is inoculated (see layout below). After growing for 18hrs at 37C, images from each of the plates were collected. Since the broth contains a cresol red indicator that changes as the pH of the media decreases, you can tell whether the part grew by noting which of the wells turned from red to yellow (as the cells grow, the media becomes more acidic).


384-deepwell test plate antibiotic quadrant layout:

1 2 3 4 ...
A Amp Cm Amp Cm
B Tet Kan Tet Kan
C Amp Cm Amp Cm
D Tet Kan Tet Kan
...



Results for each source plate:

Source Plate 1

Source Plate 2

Source Plate 3

Source Plate 4

Source Plate 5

Source Plate 6

Source Plate 7

Source Plate 8

Source Plate 9

Source Plate 10

Source Plate 11

Source Plate 12

Source Plate 13

Source Plate 14

Source Plate 15

Source Plate 16



Kit Plate Layout

Each 384-well kit plate is a composition of 4 96-well source plates. Below is a representation of which source plates went into which kit plate and in what order.

Kit Plate 1

1 2 ...
A Source Plate 1 Source Plate 2
B Source Plate 3 Source Plate 4
...



Kit Plate 2

1 2 ...
A Source Plate 5 Source Plate 6
B Source Plate 7 Source Plate 8
...



Kit Plate 3

1 2 ...
A 2006 Source Plate 1 2006 Source Plate 2
B 2006 Source Plate 3 2006 Source Plate 4
...



Kit Plate 4

1 2 ...
A Source Plate 5 (2007) Source Plate 6 (2007)
B Source Plate 7 (2007) Source Plate 8 (2007)
...