Dear Wo, The transfection method of human cells using PEI is amazing. Is this method applicable in plant tissue transformation? Besides bombardment, Agrobac. transformation, is there any new & easy technology for plant transformation? Thanks.
Jayanta ------------------------------------------------------------------------------------------- Dr. Jayanta Tarafdar Plant Pathologist & Officer in Charge AICRP on Tuber Crops (Kalyani Centre), ICAR Directorate of Research BC Krishi Viswavidyalaya Kalyani 741235 West Bengal ________________________________ From: "[email protected]" <[email protected]> To: [email protected] Sent: Wed, 27 April, 2011 10:34:24 PM Subject: Methods Digest, Vol 71, Issue 13 Send Methods mailing list submissions to [email protected] To subscribe or unsubscribe via the World Wide Web, visit http://www.bio.net/biomail/listinfo/methods or, via email, send a message with subject or body 'help' to [email protected] You can reach the person managing the list at [email protected] When replying, please edit your Subject line so it is more specific than "Re: Contents of Methods digest..." Today's Topics: 1. Re: Competitive Assay (WS) 2. Re: Copper-Glycine buffer(Ph-7.4) (WS) 3. Re: Transfection of Hi5 cells with Fugene (WS) ---------------------------------------------------------------------- Message: 1 Date: Tue, 26 Apr 2011 13:03:42 -0700 (PDT) From: WS <[email protected]> Subject: Re: Competitive Assay To: [email protected] Message-ID: <067e06b0-ca36-49c3-bb6f-29bbf3bec...@r20g2000yqd.googlegroups.com> Content-Type: text/plain; charset=ISO-8859-1 Hi Juan, if you want to have competition, you have to mix your ligands *before* you add them to your cells. If you add one of them first, it binds well to the receptor (a few socend are enough) and it will take indefinite time until an equilibrium is reached when you add the competitor. But you may use this setup for measuring turnover. Then it's called a pulse - chase experiment. Remember to add more (ligand + competitor) than you have receptors. You want competition, not complete binding. PS for a new topic, please start a new thread! Have fun! Wo On Apr 26, 3:53 pm, "Juan Li" <[email protected]> wrote: > Hello everyone, > > I would like to run a competitive assay between unlabeled ligand and > FITC-labeled ligand. How should the 2 compounds be incubated, together in a > single incubation step or one after another? What's the advantage or > disadvantage of them? > > Any suggestions? > > Thanks! > Juan ------------------------------ Message: 2 Date: Tue, 26 Apr 2011 13:04:54 -0700 (PDT) From: WS <[email protected]> Subject: Re: Copper-Glycine buffer(Ph-7.4) To: [email protected] Message-ID: <4a4c3fed-7d3b-4e89-8297-371ee94fd...@n10g2000yqf.googlegroups.com> Content-Type: text/plain; charset=ISO-8859-1 Yes Jay, you're right. I overlooked the high molarities. Wo ------------------------------ Message: 3 Date: Tue, 26 Apr 2011 14:26:25 -0700 (PDT) From: WS <[email protected]> Subject: Re: Transfection of Hi5 cells with Fugene To: [email protected] Message-ID: <[email protected]> Content-Type: text/plain; charset=ISO-8859-1 Dear Pepa, do you have access to a quicker positive control like GFP? If you don't have a fluorescence microscope, beta galactosidase or - glucuronidase and similar are suitable genes, too, as long as you can get the appropriate X-substrate. The big advantage against westerns is that you also will get an idea of the transfection efficiency, as you will see single transfected cells. Do you have the possibility to test your transfection reagent with some other cell line to check if it is ok? Are your cells healthy?any mycoplasma? really Hi5 cells? (some paranoia might be ok :) You also may dig the archives of this NG for polyethyleneimine (PEI, or simply find it here: http://www.bio.net/bionet/mm/methods/1999-December/080050.html). You only need to get the right one, it's crucial (see the protocol) and it's worth a try for sure I think, as it's horribly cheap: You'll get a whole bathtub full of final transfection reagent which is sufficient for a whole flock of sheep (see http://forums.biotechniques.com/viewtopic.php?f=18&t=13968 for details, it also works on cows), for a price much less than 1 single commercial kit). BTW, it's at least similar to JetPEI. Other hopefully useful links are in this post here: http://www.bio.net/bionet/mm/methods/2002-May/093201.html Have fun! Wo ------------------------------ _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods End of Methods Digest, Vol 71, Issue 13 *************************************** _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
