Team:Paris Bettencourt/Notebook/Drug Screening/Sunday 29th September.html
From 2013.igem.org
(Created page with "<html> <div class ="tbnote"> <h2>Drug Screening</h2> <a href="https://2013.igem.org/Team:Paris_Bettencourt/Project/Drug_Screen" target="_blank" class="tbnotelogo DSlogo"> ASDF </a...") |
|||
(3 intermediate revisions not shown) | |||
Line 11: | Line 11: | ||
</br></em></b></p> | </br></em></b></p> | ||
<!-- === Modify from here === --> | <!-- === Modify from here === --> | ||
- | Colony PCR on new Backbones. RFP and Biobrick verification primers | + | <p>Colony PCR on new Backbones. RFP and Biobrick verification primers |
This is for pSB2C and pSB3K | This is for pSB2C and pSB3K | ||
- | 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 | ||
- | Phusion PCR on Duet vectors and pSB1C3 using standard cloning primers | + | <p>Phusion PCR on Duet vectors and pSB1C3 using standard cloning primers |
- | Gel Clean up | + | <p>Gel Clean up |
- | Digestion with NcoI and AvrII | + | <p>Digestion with NcoI and AvrII |
- | Standard cloning of new backbones (1h ligation at 16C) | + | <p>Standard cloning of new backbones (1h ligation at 16C) |
- | Transform into NEBTurbo | + | <p>Transform into NEBTurbo |
- | Cutting Biobrick SirA, FprA, FdxA and Linearized backbone with EcoRI and SpeI | + | <p>Cutting Biobrick SirA, FprA, FdxA and Linearized backbone with EcoRI and SpeI |
- | Standard clonging of Biobrick parts (1h ligation at 16C) | + | <p>Standard clonging of Biobrick parts (1h ligation at 16C) |
- | Transform into NEBTurbo | + | <p>Transform into NEBTurbo |
- | Treat XbaI SpeI cut Backbone with TSAP 1h. | + | <p>Treat XbaI SpeI cut Backbone with TSAP 1h. |
- | Standard cloning of Biobrick parts (cut with XbaI, SpeI) into TSAP'd backbone (1h @ 16C) | + | <p>Standard cloning of Biobrick parts (cut with XbaI, SpeI) into TSAP'd backbone (1h @ 16C) |
+ | |||
+ | <p>Colony PCR on BL21 (DE3) dcysI pACYC SirA pET FprA FdxA and BL21 AI dcysI pACYC SirA pET FprA FdxA | ||
<!-- === To here === --> | <!-- === To here === --> | ||
</div> | </div> | ||
</html> | </html> |
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