Team:UNAM Genomics Mexico/Notebook/ANDSugar

From 2012.igem.org

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>Digested C0080 X,S. <br />
>Digested C0080 X,S. <br />
>Digested B0014 E,P [[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Digestion_protocol_.2820_.C2.B5l.29 | LIQUID CULTURE]]. <br />
>Digested B0014 E,P [[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Digestion_protocol_.2820_.C2.B5l.29 | LIQUID CULTURE]]. <br />
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<p class='captionInside'>1. 1 kb ladder <br />
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Digested B0014 with E, P and with E,X
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<h2>06/19/12</h2><br />
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>Digested B0014 with E, P and with E,X. [[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Digestion_protocol_.2820_.C2.B5l.29 | LIQUID CULTURE]]. <br />
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>Ran gels with yesterday’s digestions [[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Gel_electrophoresis | GEL ELECTROPHORESIS PROTOCOL]] . <br />
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2 B0014 E,X and 3 B0014 E,P were discarded because of problems in the preparation. <br />
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C0080 is in the first lane. <br />
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>Extracted from gel C0080 X,S [[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Gel_extraction_protocol | LIQUID CULTURE]]. <br />
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>Made glycerols from 4 plates LB Km DH5α pSB2K3 [[Team:UNAM_Genomics_Mexico/Notebook/Protocols#GLYCEROL_PROTOCOL | GLYCEROL PROTOCOL]] . <br />
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>Take out AmyE 5’ from distribution [DNA KIT PLATE INSTRUCTIONS]. <br />
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AmyE 5’ 18K plate 3 2010, 2011, 2012<br />
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AmyE 3’ 18M plate 3 2010, 2011, 2012<br />
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>Digested B0014 with E,X and E,P again [[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Digestion_protocol_.2820_.C2.B5l.29 | LIQUID CULTURE]]. <br />
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<img src='https://static.igem.org/mediawiki/2012/7/73/UnamgenomicsandsugarBitacora_7.jpg' height="300"/>
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<div class='captionaqua'>
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<p class='captionInside'>1.B0014 E,X<br />
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2.B0014 E,P<br />
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3. ΩSpr/Strr  E<br />
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4.C0080 X,S<br />
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5.E1010 X,S<br />
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6.2 μl  ladder<br />
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06/22/12<br />
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>Ran gels with digestions B0014 with E,X and E,P [[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Gel_electrophoresis | GEL ELECTROPHORESIS PROTOCOL]]. <br />
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>Dephosphated B0014 E,X and B0014 E,P [DESPHOPHORYLATION PROTOCOL]. <br />
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Revision as of 05:26, 23 September 2012


UNAM-Genomics_Mexico


Contents

Arabinose/Xylose AND Gate



Nanotubes!! The logic Random info




  • 1. 1 kb ladder
    2.E1010



06/07/12


PY BROTH 10g salt/L
PSB2K3 kanamicin
We transformed RFP E1010.
plate 1 18F
plate 2 17E
Stock 50 mg/mL
E.coli 1/1000
TRANSFORMATION PROTOCOL.
We made liquid cultures with colonies LIQUID CULTURE.
We extracted plasmids PLASMID EXTRACTION PROTOCOL.
We ran a gel to check the extraction GEL ELECTROPHORESIS PROTOCOL.




06/12/12


We made glycerols of the bacteria
GLYCEROL PROTOCOL .
We transformed plasmid pHp45 TRANSFORMATION PROTOCOL.

  • 1. 1 kb ladder
    2.E1010



06/13/12


We ran gel and extracted from gel LIQUID CULTURE.
We made liquid cultures LIQUID CULTURE and lysed [LYSIS PROTOCOL].














  • 1. 1 kb ladder
    We extracted from gel



06/14/12


We extracted from gel LIQUID CULTURE.
We made glycerols of pHp45 Ω GLYCEROL PROTOCOL .
We transformed PSB2K3 Km (Kanamycin) 5C BBa_J04450 PSB4A5 Am (ampicillin) 1I BBa_J04450 AraC BBa_C0080
2012 14L plate 1 pSB2K3 Km+
2011 14L plate 1 pSB2K3 Km+
2012 14L plate 1 pSB2K3 Km+
TRANSFORMATION PROTOCOL.





  • 1. 1 kb ladder
    pHp45 Ω with E/P
    We extracted from gel



06/15/12


> Digested pHp45 Ω with E/P LIQUID CULTURE.
> We extracted plasmid with kit.
> Ran a gel GEL ELECTROPHORESIS PROTOCOL . (5)
>Extracted from gel LIQUID CULTURE.

ARAC
>From the overnight plate we prepared liquid culture LIQUID CULTURE.
>We left them incubating overnight.
>We left a plate (LB Km DH5α C0080 and a control).





06/18/12


>Plasmid extraction AraC with kit.
>C0080-psB2K3 915 bp
>Streaked 4 LB Km 30 plates DH5α psB2K3.
>It seemed as though there were two different colonies (white) and there had been a mutation. Normally they are red.
>Ran gel with Spr/Strr (eppendorf -20ºC) GEL ELECTROPHORESIS PROTOCOL.
>Made glycerols from one tube of DH5α C0080 Km 50 -> 04.
>Made glycerols from one tube of DH5α PSB4A5 Amp 100 -> 05 GLYCEROL PROTOCOL.
>Digested C0080 X,S.
>Digested B0014 E,P LIQUID CULTURE.


  • 1. 1 kb ladder
    Digested B0014 with E, P and with E,X





06/19/12


>Digested B0014 with E, P and with E,X. LIQUID CULTURE.
>Ran gels with yesterday’s digestions GEL ELECTROPHORESIS PROTOCOL .
2 B0014 E,X and 3 B0014 E,P were discarded because of problems in the preparation.
C0080 is in the first lane.
>Extracted from gel C0080 X,S LIQUID CULTURE.
>Made glycerols from 4 plates LB Km DH5α pSB2K3 GLYCEROL PROTOCOL .
>Take out AmyE 5’ from distribution [DNA KIT PLATE INSTRUCTIONS].
AmyE 5’ 18K plate 3 2010, 2011, 2012
AmyE 3’ 18M plate 3 2010, 2011, 2012
>Digested B0014 with E,X and E,P again LIQUID CULTURE.






  • 1.B0014 E,X
    2.B0014 E,P
    3. ΩSpr/Strr E
    4.C0080 X,S
    5.E1010 X,S
    6.2 μl ladder



06/22/12
>Ran gels with digestions B0014 with E,X and E,P GEL ELECTROPHORESIS PROTOCOL.

>Dephosphated B0014 E,X and B0014 E,P [DESPHOPHORYLATION PROTOCOL].

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