Team:UNICAMP-EMSE Brazil/Notebook/25 August 2011

From 2011.igem.org

(Difference between revisions)
(→Digestions recipe)
 
(6 intermediate revisions not shown)
Line 3: Line 3:
[[Image:UNICAMP-EMSE Brazil Gel1 25 08 11.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 1, 25/08/2011.'''</font> <br>Gel 1:RBS+TolC+T C1 (d)/ RBS+TolC+T C1 (nd)/ RBS+TolC+T C2 (d)/ RBS+TolC+T C2 (nd)/ RBS+TolC+T C3 (d)/ RBS+TolC+T C3 (nd)/ RBS+TolC+T C4 (d)/ RBS+TolC+T C4 (nd).]]
[[Image:UNICAMP-EMSE Brazil Gel1 25 08 11.jpg|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 1, 25/08/2011.'''</font> <br>Gel 1:RBS+TolC+T C1 (d)/ RBS+TolC+T C1 (nd)/ RBS+TolC+T C2 (d)/ RBS+TolC+T C2 (nd)/ RBS+TolC+T C3 (d)/ RBS+TolC+T C3 (nd)/ RBS+TolC+T C4 (d)/ RBS+TolC+T C4 (nd).]]
==25 August 2011==
==25 August 2011==
-
===RBS+TolC+T and RBS+SocR_T===
+
===RBS+TolC+T and RBS+SoxR_T===
'''Objective:'''   
'''Objective:'''   
:*Checking “RBS+TolC+T” and “RBS+SoxR+T” purified by Mini-prep and digested with E-P.
:*Checking “RBS+TolC+T” and “RBS+SoxR+T” purified by Mini-prep and digested with E-P.
Line 43: Line 43:
<br>
<br>
<br>
<br>
 +
 +
===Isolate digested biobricks from their vectors===
 +
[[Image:UNICAMP-EMSE_Brazil_Gel3_25_08_11.png|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 3, 25/08/2011.'''</font>]]
 +
*'''Ladder:'''
 +
:*Bio-Rad 100bp – 10.000bp
 +
'''Objective:''' 
 +
:*isolate digested biobricks from their vector  (volume of ladder applied 6uL; digestion sample:40uL digested sample+4uL buffer)
 +
*'''Samples: '''
 +
:*1: RBS+TolC+terminator (digested) ;
 +
:*2: RBS+TolC+terminator (non-digested);
 +
:*3: RBS+IL10 (digested);
 +
:*4: RBS+IL10 (non-digested);
 +
:*5: RBS+IL12 (digested);
 +
6 :RBS+IL12 (Non-digested); 
 +
:*7: RBS+SoxR+Terminator (digested);
 +
:*8: RBS+SoxR+Terminator (non-digested).
 +
:*9: RBS+SoxR+Terminator (digested);
 +
 +
<br clear="all" />
 +
*'''<font color=teal>Results:'''</font>
 +
<font align=center>
 +
{|border="1" cellpadding="5" cellspacing="0" align="center"
 +
|-
 +
! scope="col" style="background:#efefef;" | Gene
 +
! scope="col" style="background:#efefef;" | Total size (gene+vector)
 +
! scope="col" style="background:#efefef;" | Linear vector size
 +
! scope="col" style="background:#efefef;" | Gene size
 +
! scope="col" style="background:#efefef;" | Result
 +
|-
 +
|'''RBS+TolC+terminator'''
 +
|4880 pb
 +
|3300 pb
 +
|1675 pb
 +
|Bright higher band
 +
|-
 +
|''' RBS+IL10'''
 +
|3345 pb
 +
|2800 pb
 +
|545 pb
 +
|Didn’t work
 +
|-
 +
|'''RBS+IL12'''
 +
|4458 pb
 +
|2800 pb
 +
|1658 pb
 +
|OK
 +
|-
 +
|'''RBS+SoxR+Terminator'''
 +
|~3823 pb
 +
|3300 pb
 +
|1618 pb
 +
|OK
 +
|-
 +
|'''RBS+SoxR+Terminator'''
 +
|~3823 pb
 +
|3300 pb
 +
|1618 pb
 +
|OK
 +
|}</font>
 +
<br>
 +
<br>
 +
 +
[[Image:UNICAMP-EMSE_Brazil_Gel4_25_08_11.png|thumb|right|300px|<font color: blue>'''Electrophoresis Gel 4, 25/08/2011.'''</font>]]
 +
*'''Ladder:'''
 +
:*Bio-Rad 100bp – 10.000bp
 +
'''Objective:''' 
 +
:*quantification of the purified bands from the previous electrophoresis
 +
*'''Samples: '''
 +
:*1: RBS+TolC+terminator ; 
 +
:*2: RBS+IL12 ;
 +
:*3: RBS+SoxR+Terminator
 +
 +
*'''<font color=teal>Results:'''</font>
 +
*Their concentration seems quite good!
 +
<br>
 +
<br>
 +
<br>
 +
 +
===Digestions recipes===
 +
*'''RBS+TolC+Terminator (E-X)'''
 +
:*29ul  - RBS+TolC+Terminator
 +
:*8ul    - 10X Tango Buffer
 +
:*1ul    - EcoRI
 +
:*2ul  - XbaI
 +
:*'''TOTAL = 40ul'''
 +
 +
*'''RBS+IL-10 (X-P)'''
 +
:*32ul  - RBS+IL-10
 +
:*4ul    - 10X Tango Buffer
 +
:*1.4ul  - XbaI
 +
:*2.6ul  - PstI
 +
:*'''TOTAL = 40ul'''
 +
 +
*'''RBS+IL-12 (X-P)'''
 +
:*32ul  - RBS+IL-12
 +
:*4ul    - 10X Tango Buffer
 +
:*1.4ul  - XbaI
 +
:*2.6ul  - PstI
 +
:*'''TOTAL = 40ul'''
 +
 +
*'''RBS+SoxR (X-P)'''
 +
:*32ul  - RBS+SoxR
 +
:*4ul    - 10X Tango Buffer
 +
:*1.4ul  - XbaI
 +
:*2.6ul  - PstI
 +
:*'''TOTAL = 40ul'''
 +
 +
 +
'''Next Tasks:'''
 +
#Incubate reactions at 37°C for 3-4hours.
 +
#Electrophoresis (loading the whole digestions volume - join two or more wells in the gel)
 +
#Purify the bands
 +
#Electrophoresis to confirm (load 5ul of gel purified digestion)
 +
 +
 +
<html>
 +
<img src="https://static.igem.org/mediawiki/2011/c/cb/UNICAMP-EMSE_Brazil_Stresswars.png" width="893px"></img>
 +
</html>
 +
 +
 +
<html>
 +
<head>
 +
<script language='JavaScript' type='text/JavaScript'>
 +
 +
function makeQuote() {
 +
 +
Q = new Array();
 +
 +
 +
Q[0] = "<i>I’m Luke Skywalker, I’m here to rescue you.</i>Luke, Star Wars: Episode IV-A New Hope (1977)"
 +
Q[1] = "<i>Watch your mouth kid, or you’ll find yourself floating home.</i>Star Wars: Episode IV-A New Hope (1977)"
 +
Q[2] = "<i>Evacuate in our moment of triumph? I think you overestimate their chances.</i> Star Wars: Episode IV-A New Hope (1977)"
 +
Q[3] = "<i>The Force is strong with this one.</i> Star Wars: Episode IV-A New Hope (1977)"
 +
Q[4] = "<i> I have you now!</i> Darth Vader, Star Wars: Episode IV-A New Hope (1977)"
 +
Q[5] = "<i> Fear is the path to the dark side. Fear leads to anger, anger leads to hate, hate leads to suffering.</i> MsC Yoda, Star Wars Episode I: The Phantom Menace (1999)"
 +
Q[6] = "<i> No blasters! No blasters!.</i> A bartender, Star Wars: Episode IV-A New Hope (1977)"
 +
Q[7] = "<i> Go to the center of the gravity's pull, and find your planet you will...</i> M.Sc. Yoda, Star Wars Episode II: Attack of the Clones (2002)"
 +
Q[8] = "<i> Clear, your mind must be if you are to discover the real villains behind the plot.</i> M.Sc. Yoda, Star Wars Episode II: Attack of the Clones (2002)"
 +
Q[9] = "<i> Much to learn, you still have.</i> M.Sc. Yoda, Star Wars Episode II: Attack of the Clones (2002)"
 +
Q[10] = "<i> Meditate on this, I will.</i> M.Sc. Yoda, Star Wars Episode II: Attack of the Clones (2002)"
 +
Q[11] = "<i> Use the Force, Luke.</i> M.Sc. Yoda, Star Wars Episode II: Attack of the Clones (2002)"
 +
Q[12] = "<i> May the Force be with you.</i> Mentioned several times throughout the Star Wars saga"
 +
Q[13] = "<i> I really need a Jedi Bacteria.</i> Iolanda Albuquerque, UNICAMP-EMSE team advisor"
 +
Q[14] = "<i> Another splendid day in Brazil!!!</i> Marc Emery, UNICAMP-EMSE team student"
 +
Q[15] = "<i> Dont' cut, let your hair volume expressing all their feelings...</i> Thibault Sabattier (UNICAMP-EMSE team student) about Nemailla's hair"
 +
Q[16] = "<i> What a good life!!</i> Louise Marais (UNICAMP-EMSE team student) about living in Brazil"
 +
Q[17] = "<i> Gosh!I will have nigthmares tonight! </i> Danieli Gonçalves (UNICAMP-EMSE team student) about Thibault wearing Marianna's dress"
 +
Q[18] = "<i> I will revenge myself against you and Marc Emery!!!</i> Nemailla Bonturi (UNICAMP-EMSE team advisor)"
 +
Q[19] = "<i> Yeah, those dresses deserve us!</i> Marc Emery (UNICAMP-EMSE team student) about Marianna's dresses"
 +
Q[20] = "<i> I am really starting getting worried with you guys!!</i> Iolanda Albuquerque (UNICAMP-EMSE team advisor)"
 +
 +
 +
 +
 +
 +
 +
 +
 +
 +
index = Math.floor(Math.random() * Q.length);
 +
 +
document.writeln(Q[index]);
 +
 +
}
 +
 +
</script>
 +
 +
</head>
 +
 +
<style>
 +
#newquote {color:gray; font-family:verdana; font-size:11px; text-align:right; padding:5px;}
 +
 +
#newquote i {display:block; color:black; font-family:georgia, trebuchet ms; font-size:13px;}
 +
 +
#newquote small {display:block; text-align:right; margin-top:10px;}
 +
 +
]]></b:skin>
 +
</style>
 +
</html>
 +
 +
 +
<html>
 +
<head>
 +
<div id='newquote'>
 +
 +
<script language="javascript">
 +
 +
makeQuote();
 +
 +
</script>
 +
</div>
 +
</head>
 +
</html>

Latest revision as of 00:17, 29 September 2011

Home Project Methods Results Data Team Notebook Human Practices Safety Profile Sponsors Wix

Click here to see our PROTOCOLS

Contents

Notebook

Click on a date to see what we have done!

May
MTWTFSS
            [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/1_May_2011&action=edit 1]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/2_May_2011&action=edit 2] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/3_May_2011&action=edit 3] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/4_May_2011&action=edit 4] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/5_May_2011&action=edit 5] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/6_May_2011&action=edit 6] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/7_May_2011&action=edit 7] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/8_May_2011&action=edit 8]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/9_May_2011&action=edit 9] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/10_May_2011&action=edit 10] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/11_May_2011&action=edit 11] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/12_May_2011&action=edit 12] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/13_May_2011&action=edit 13] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/14_May_2011&action=edit 14] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/15_May_2011&action=edit 15]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/16_May_2011&action=edit 16] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/17_May_2011&action=edit 17] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/18_May_2011&action=edit 18] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/19_May_2011 19] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/20_May_2011&action=edit 20] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/21_May_2011&action=edit 21] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/22_May_2011&action=edit 22]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/23_May_2011&action=edit 23] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/24_May_2011&action=edit 24] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/25_May_2011 25] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/26_May_2011 26] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/27_May_2011 27] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/28_May_2011&action=edit 28] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/29_May_2011&action=edit 29]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/30_May_2011 30] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/31_May_2011 31]
June
MTWTFSS
    [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/1_June_2011 1] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/2_June_2011 2] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/3_June_2011 3] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/4_June_2011&action=edit 4] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/5_June_2011&action=edit 5]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/6_June_2011 6] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/7_June_2011 7] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/8_June_2011 8] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/9_June_2011 9] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/10_June_2011 10] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/11_June_2011&action=edit 11] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/12_June_2011&action=edit 12]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/13_June_2011 13] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/14_June_2011&action=edit 14] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/15_June_2011 15] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/16_June_2011 16] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/17_June_2011 17] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/18_June_2011&action=edit 18] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/19_June_2011&action=edit 19]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/20_June_2011 20] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/21_June_2011 21] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/22_June_2011 22] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/23_June_2011&action=edit 23] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/24_June_2011&action=edit 24] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/25_June_2011&action=edit 25] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/26_June_2011&action=edit 26]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/27_June_2011&action=edit 27] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/28_June_2011&action=edit 28] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/29_June_2011 29] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/30_June_2011&action=edit 30]
July
MTWTFSS
        [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/1_July_2011&action=edit 1] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/2_July_2011&action=edit 2] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/3_July_2011 3]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/4_July_2011&action=edit 4] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/5_July_2011&action=edit 5] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/6_July_2011&action=edit 6] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/7_July_2011 7] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/8_July_2011 8] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/9_July_2011&action=edit 9] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/10_July_2011&action=edit 10]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/11_July_2011 11] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/12_July_2011 12] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/13_July_2011 13] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/14_July_2011 14] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/15_July_2011 15] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/16_July_2011 16] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/17_July_2011&action=edit 17]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/18_July_2011 18] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/19_July_2011 19] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/20_July_2011&action=edit 20] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/21_July_2011 21] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/22_July_2011 22] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/23_July_2011&action=edit 23] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/24_July_2011&action=edit 24]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/25_July_2011 25] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/26_July_2011 26] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/27_July_2011 27] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/28_July_2011 28] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/29_July_2011&action=edit 29] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/30_July_2011&action=edit 30] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/31_July_2011&action=edit 31]
August
MTWTFSS
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/1_August_2011 1] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/2_August_2011 2] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/3_August_2011&action=edit 3] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/4_August_2011&action=edit 4] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/5_August_2011&action=edit 5] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/6_August_2011&action=edit 6] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/7_August_2011&action=edit 7]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/8_August_2011&action=edit 8] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/9_August_2011&action=edit 9] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/10_August_2011&action=edit 10] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/11_August_2011&action=edit 11] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/12_August_2011&action=edit 12] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/13_August_2011&action=edit 13] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/14_August_2011&action=edit 14]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/15_August_2011&action=edit 15] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/16_August_2011&action=edit 16] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/17_August_2011&action=edit 17] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/18_August_2011&action=edit 18] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/19_August_2011 19] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/20_August_2011 20] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/21_August_2011 21]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/22_August_2011 22] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/23_August_2011 23] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/24_August_2011 24] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/25_August_2011 25] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/26_August_2011 26] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/27_August_2011 27] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/28_August_2011&action=edit 28]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/29_August_2011 29] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/30_August_2011 30] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/31_August_2011 31]
September
MTWTFSS
      [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/1_September_2011 1] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/2_September_2011 2] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/3_September_2011&action=edit 3] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/4_September_2011&action=edit 4]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/5_September_2011 5] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/6_September_2011 6] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/7_September_2011 7] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/8_September_2011 8] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/9_September_2011 9] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/10_September_2011 10] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/11_September_2011 11]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/12_September_2011 12] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/13_September_2011 13] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/14_September_2011 14] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/15_September_2011&action=edit 15] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/16_September_2011 16] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/17_September_2011 17] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/18_September_2011&action=edit 18]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/19_September_2011 19] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/20_September_2011&action=edit 20] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/21_September_2011 21] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/22_September_2011 22] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/23_September_2011 23] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/24_September_2011 24] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/25_September_2011 25]
[http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/26_September_2011 26] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/27_September_2011 27] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/28_September_2011 28] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/29_September_2011 29] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/30_September_2011&action=edit 30]
October
MTWTFSS
          [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/1_October_2011&action=edit 1] [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/2_October_2011 2]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/3_October_2011&action=edit 3] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/4_October_2011&action=edit 4] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/5_October_2011&action=edit 5] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/6_October_2011&action=edit 6] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/7_October_2011&action=edit 7] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/8_October_2011&action=edit 8] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/9_October_2011&action=edit 9]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/10_October_2011&action=edit 10] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/11_October_2011&action=edit 11] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/12_October_2011&action=edit 12] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/13_October_2011&action=edit 13] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/14_October_2011&action=edit 14] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/15_October_2011&action=edit 15] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/16_October_2011&action=edit 16]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/17_October_2011&action=edit 17] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/18_October_2011&action=edit 18] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/19_October_2011&action=edit 19] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/20_October_2011&action=edit 20] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/21_October_2011&action=edit 21] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/22_October_2011&action=edit 22] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/23_October_2011&action=edit 23]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/24_October_2011&action=edit 24] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/25_October_2011&action=edit 25] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/26_October_2011&action=edit 26] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/27_October_2011&action=edit 27] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/28_October_2011&action=edit 28] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/29_October_2011&action=edit 29] [http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/30_October_2011&action=edit 30]
[http://2011.igem.org/wiki/index.php?title=Team:UNICAMP-EMSE_Brazil/Notebook/31_October_2011&action=edit 31]


Electrophoresis Gel 1, 25/08/2011.
Gel 1:RBS+TolC+T C1 (d)/ RBS+TolC+T C1 (nd)/ RBS+TolC+T C2 (d)/ RBS+TolC+T C2 (nd)/ RBS+TolC+T C3 (d)/ RBS+TolC+T C3 (nd)/ RBS+TolC+T C4 (d)/ RBS+TolC+T C4 (nd).

25 August 2011

RBS+TolC+T and RBS+SoxR_T

Objective:

  • Checking “RBS+TolC+T” and “RBS+SoxR+T” purified by Mini-prep and digested with E-P.
Electrophoresis Gel 2, 25/08/2011.
RBS+SoxR+T C1 (d)/ RBS+SoxR+T C1 (nd)/ RBS+SoxR+T C2 (d)/ RBS+SoxR+T C2 (nd)/ RBS+SoxR+T C3 (d)/ RBS+SoxR+T C3 (nd)/ RBS+SoxR+T C4 (d)/ RBS+SoxR+T C4 (nd).
  • Ladder:
  • Bio-Rad 100bp – 10.000bp
  • Samples:
  • Gel 1: RBS+TolC+T C1 (d)/ RBS+TolC+T C1 (nd)/ RBS+TolC+T C2 (d)/ RBS+TolC+T C2 (nd)/ RBS+TolC+T C3 (d)/ RBS+TolC+T C3 (nd)/ RBS+TolC+T C4 (d)/ RBS+TolC+T C4 (nd)
  • Gel 2: RBS+SoxR+T C1 (d)/ RBS+SoxR+T C1 (nd)/ RBS+SoxR+T C2 (d)/ RBS+SoxR+T C2 (nd)/ RBS+SoxR+T C3 (d)/ RBS+SoxR+T C3 (nd)/ RBS+SoxR+T C4 (d)/ RBS+SoxR+T C4 (nd)

OBS:

  • D=digested
  • Nd= non-digested


  • Results:

Gene Total size (gene+vector) Linear vector size Gene size Result
RBS+TolC+T ~4880 pb 3300 pb 1675 pb OK: complete digestion
RBS+SoxR+T ~3823 pb 3300 pb 618 pb Only C4 did not digested.
C1 and C3 has a gene fragment near 500 pb
and C2 has one near 700 bp.
Wich one is the correct?



Isolate digested biobricks from their vectors

Electrophoresis Gel 3, 25/08/2011.
  • Ladder:
  • Bio-Rad 100bp – 10.000bp

Objective:

  • isolate digested biobricks from their vector (volume of ladder applied 6uL; digestion sample:40uL digested sample+4uL buffer)
  • Samples:
  • 1: RBS+TolC+terminator (digested) ;
  • 2: RBS+TolC+terminator (non-digested);
  • 3: RBS+IL10 (digested);
  • 4: RBS+IL10 (non-digested);
  • 5: RBS+IL12 (digested);

6 :RBS+IL12 (Non-digested);

  • 7: RBS+SoxR+Terminator (digested);
  • 8: RBS+SoxR+Terminator (non-digested).
  • 9: RBS+SoxR+Terminator (digested);


  • Results:

Gene Total size (gene+vector) Linear vector size Gene size Result
RBS+TolC+terminator 4880 pb 3300 pb 1675 pb Bright higher band
RBS+IL10 3345 pb 2800 pb 545 pb Didn’t work
RBS+IL12 4458 pb 2800 pb 1658 pb OK
RBS+SoxR+Terminator ~3823 pb 3300 pb 1618 pb OK
RBS+SoxR+Terminator ~3823 pb 3300 pb 1618 pb OK



Electrophoresis Gel 4, 25/08/2011.
  • Ladder:
  • Bio-Rad 100bp – 10.000bp

Objective:

  • quantification of the purified bands from the previous electrophoresis
  • Samples:
  • 1: RBS+TolC+terminator ;
  • 2: RBS+IL12 ;
  • 3: RBS+SoxR+Terminator
  • Results:
  • Their concentration seems quite good!




Digestions recipes

  • RBS+TolC+Terminator (E-X)
  • 29ul - RBS+TolC+Terminator
  • 8ul - 10X Tango Buffer
  • 1ul - EcoRI
  • 2ul - XbaI
  • TOTAL = 40ul
  • RBS+IL-10 (X-P)
  • 32ul - RBS+IL-10
  • 4ul - 10X Tango Buffer
  • 1.4ul - XbaI
  • 2.6ul - PstI
  • TOTAL = 40ul
  • RBS+IL-12 (X-P)
  • 32ul - RBS+IL-12
  • 4ul - 10X Tango Buffer
  • 1.4ul - XbaI
  • 2.6ul - PstI
  • TOTAL = 40ul
  • RBS+SoxR (X-P)
  • 32ul - RBS+SoxR
  • 4ul - 10X Tango Buffer
  • 1.4ul - XbaI
  • 2.6ul - PstI
  • TOTAL = 40ul


Next Tasks:

  1. Incubate reactions at 37°C for 3-4hours.
  2. Electrophoresis (loading the whole digestions volume - join two or more wells in the gel)
  3. Purify the bands
  4. Electrophoresis to confirm (load 5ul of gel purified digestion)




Retrieved from "http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Notebook/25_August_2011"