Team:Paris Bettencourt/Notebook/Drug Screening/Sunday 29th September.html

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Colony PCR on new Backbones.  RFP and Biobrick verification primers
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<p>Colony PCR on new Backbones.  RFP and Biobrick verification primers
This is for pSB2C and pSB3K
This is for pSB2C and pSB3K
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Colony PCR on SirA, FprA, and FdxA Biobricked parts
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<p>Colony PCR on SirA, FprA, and FdxA Biobricked parts
Using Biobrick primers and SirA, FprA and FdxA primers respectfully
Using Biobrick primers and SirA, FprA and FdxA primers respectfully
 +
https://2013.igem.org/File:20130929_BioB.jpg
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https://2013.igem.org/File:20130929_RFP.jpg
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https://2013.igem.org/File:20190929_pSB3K.jpg
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Phusion PCR on Duet vectors and pSB1C3 using standard cloning primers
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<p>Phusion PCR on Duet vectors and pSB1C3 using standard cloning primers
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Gel Clean up
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<p>Gel Clean up
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Digestion with NcoI and AvrII
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<p>Digestion with NcoI and AvrII
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Standard cloning of new backbones (1h ligation at 16C)
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<p>Standard cloning of new backbones (1h ligation at 16C)
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Transform into NEBTurbo
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<p>Transform into NEBTurbo
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Cutting Biobrick SirA, FprA, FdxA and Linearized backbone with EcoRI and SpeI
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<p>Cutting Biobrick SirA, FprA, FdxA and Linearized backbone with EcoRI and SpeI
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Standard clonging of Biobrick parts (1h ligation at 16C)
+
<p>Standard clonging of Biobrick parts (1h ligation at 16C)
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Transform into NEBTurbo
+
<p>Transform into NEBTurbo
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Treat XbaI SpeI cut Backbone with TSAP 1h.
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<p>Treat XbaI SpeI cut Backbone with TSAP 1h.
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Standard cloning of Biobrick parts (cut with XbaI, SpeI) into TSAP'd backbone (1h @ 16C)
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<p>Standard cloning of Biobrick parts (cut with XbaI, SpeI) into TSAP'd backbone (1h @ 16C)
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<p>Colony PCR on BL21 (DE3) dcysI pACYC SirA pET FprA FdxA and BL21 AI dcysI pACYC SirA pET FprA FdxA
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Latest revision as of 20:03, 29 September 2013

Drug Screening

Sunday 29th September

PCR's and more!

Colony PCR on new Backbones. RFP and Biobrick verification primers This is for pSB2C and pSB3K

Colony PCR on SirA, FprA, and FdxA Biobricked parts Using Biobrick primers and SirA, FprA and FdxA primers respectfully https://2013.igem.org/File:20130929_BioB.jpg https://2013.igem.org/File:20130929_RFP.jpg https://2013.igem.org/File:20190929_pSB3K.jpg

Phusion PCR on Duet vectors and pSB1C3 using standard cloning primers

Gel Clean up

Digestion with NcoI and AvrII

Standard cloning of new backbones (1h ligation at 16C)

Transform into NEBTurbo

Cutting Biobrick SirA, FprA, FdxA and Linearized backbone with EcoRI and SpeI

Standard clonging of Biobrick parts (1h ligation at 16C)

Transform into NEBTurbo

Treat XbaI SpeI cut Backbone with TSAP 1h.

Standard cloning of Biobrick parts (cut with XbaI, SpeI) into TSAP'd backbone (1h @ 16C)

Colony PCR on BL21 (DE3) dcysI pACYC SirA pET FprA FdxA and BL21 AI dcysI pACYC SirA pET FprA FdxA