Team:Debrecen-Hungary/protocols
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=== Cell Passaging === | === Cell Passaging === | ||
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+ | Cell passaging or splitting is a technique that enables an individual to keep cells alive and growing under cultured conditions for extended periods of time. Cells should be passed when they are 90%-100% confluent. You have to do the cell passage on every second to fourth day (i.e. on every Monday, Wednesday and Friday). | ||
+ | After reaching the confluency, the cells do not get enough nutrients and do not have enough place where they can extend. The colour of the medium switches from reddish-pink to orange or yellow which shows acidic metabolic products. | ||
+ | [http://www.openwetware.org/wiki/IGEM:University_of_Debrecen:Cell_Passaging Read more...] | ||
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=== Media PEI Preparation === | === Media PEI Preparation === | ||
=== Transfection === | === Transfection === |
Revision as of 15:26, 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 electrophoresis
Notebook protocols utilized by the tissue culture subteamCell PassagingCell passaging or splitting is a technique that enables an individual to keep cells alive and growing under cultured conditions for extended periods of time. Cells should be passed when they are 90%-100% confluent. You have to do the cell passage on every second to fourth day (i.e. on every Monday, Wednesday and Friday). After reaching the confluency, the cells do not get enough nutrients and do not have enough place where they can extend. The colour of the medium switches from reddish-pink to orange or yellow which shows acidic metabolic products. Read more... Media PEI PreparationTransfectionLigand TreatmentNotebook protocols utilized by the Luciferase subteamMeasuring Luciferase activity with the Victor plate reader |