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Team:Paris Bettencourt/Notebook/Drug Screening/Sunday 29th September.html
From 2013.igem.org
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<p>Colony PCR on SirA, FprA, and FdxA Biobricked parts | <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 | ||
| + | https://2013.igem.org/File:20130929_RFP.jpg | ||
| + | https://2013.igem.org/File:20190929_pSB3K.jpg | ||
<p>Phusion PCR on Duet vectors and pSB1C3 using standard cloning primers | <p>Phusion PCR on Duet vectors and pSB1C3 using standard cloning primers | ||
Latest revision as of 20:03, 29 September 2013
Drug Screening
ASDFSunday 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
