"
Team:Debrecen-Hungary/protocols
From 2010.igem.org
Ophir Keret (Talk | contribs) (→Restriction enzyme digestion) |
Ophir Keret (Talk | contribs) (→PCR product purification) |
||
| Line 49: | Line 49: | ||
[http://openwetware.org/wiki/IGEM:University_of_Debrecen:Restriction_biobrick_parts#Notes Read more...] | [http://openwetware.org/wiki/IGEM:University_of_Debrecen:Restriction_biobrick_parts#Notes Read more...] | ||
| - | === PCR product purification === | + | === PCR product purification / Gel purification === |
| + | |||
| + | [http://www.openwetware.org/wiki/IGEM:University_of_Debrecen:PCR_purification_from_Agarose_Gel Read more...] | ||
| + | |||
=== Gel electrophoresis === | === Gel electrophoresis === | ||
=== Gel purification === | === Gel purification === | ||
Revision as of 15:24, 28 September 2010
Welcome To Our Notebook"Science is nothing else but the art of proper reproducibility. "The iGEM experience is not merely a project or a conference, but it was the way that most of our students got acquainted with the world of biological research laboratory. Pipettes, solutions, gels, electrodes, dishes and other scary machinery quickly filled our lives. From day one we saw the vast importance of teaching our students to keep a proper laboratory journal. As time passed and our project grew., and with growth sprouted the idea of keeping an electronic laboratory journal with texts and video’s depicting the proper way of doing our niche of science. And so, we present to you our combined effort a text and video version ContentsNotebook protocols utilized by the bacterial work subteam Making Lurea's Broth - Transformation of competent cells - Mini Prep Notebook protocols utilized by the molecular tools subteam Restriction enzyme digestion - PCR Purification - Gel electrophoresis - Gel purification Notebook protocols utilized by the tissue culture subteam Cell Passaging - Media PEI Preparation - Transfection - Ligand Treatment Notebook protocols utilized by the Luciferase subteam Measuring Luciferase activity with the Victor plate reader Notebook protocols utilized by the bacterial work subteamMaking Lurea's BrothTransformation of competent cellsTransformation is the process of introducing foreign DNA (e.g plasmids, BAC) into a bacterium. Bacterial cells into which foreign DNA can be transformed are called competent. Some bacteria are naturally competent (e.g B. subtilis), whereas others such as E. coli are not naturally competent. Non-competent cells can be made competent and then transformed via one of two main approaches; chemical transformation and electroporation. It is important to note we have tested transformations of the distribution kit with this protocol.Read more... Mini PrepThis protocol is designed for purification of up to 20 μg of high-copy plasmid DNA from 1–5 ml overnight cultures of E. coli in LB (Luria-Bertani) medium Read more... Notebook protocols utilized by the molecular tools subteamRestriction enzyme digestionBioBrick standard biological parts are flanked by well characterized upstream and downstream sequences which are technically not considered part of the BioBrick part (aka prefix and suffix). These up/down stream segments contain restriction sites for specific restriction enzymes, which allows for the simple creation of larger BioBrick parts by chaining together smaller ones in any desired order. Read more... PCR product purification / Gel purificationGel electrophoresisGel purificationNotebook protocols utilized by the tissue culture subteamCell PassagingMedia PEI PreparationTransfectionLigand TreatmentNotebook protocols utilized by the Luciferase subteamMeasuring Luciferase activity with the Victor plate reader |
