"High background in Amino-allyl labelling "

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masako Aug-21-01, 05:01 PM (PDT) "High background in Amino-allyl labelling " Hi, I have been doing some microarry work and I have been using Cy3/5-dCTP and they are working pretty well follwing basicly Stanford protocol with a few changes. Now I have tried to use Amino-allyl labelling and I have done two hyb. Both of them gave me very strong background and black spots. I am using Cot1 DNA, salmon sperm DNA and Poly A in formamide based buffer as many of the protocols say. Does anyone know why this may be the case or any word of advice on amino-allyl labelling will be much appreciated. Thanks Masako   Alert Edit | Reply | Reply With Quote | Top

Table of contents

RE: High background in Amino-allyl ..., Chantal, Aug-23-01, (1) RE: High background in Amino-allyl ..., masako, Aug-26-01, (2) RE: High background in Amino-allyl ..., zhengd, Mar-27-02, (3) RE: High background in Amino-allyl ..., cyj, Apr-04-02, (4) RE: High background in Amino-allyl ..., marchini, Apr-13-02, (5) RE: High background in Amino-allyl ..., mojgana, Apr-16-02, (6) RE: High background in Amino-allyl ..., Chantal, Apr-18-02, (7) RE: High background in Amino-allyl ..., marchini, Apr-21-02, (8) RE: High background in Amino-allyl ..., alli, Apr-23-02, (9) RE: High background in Amino-allyl ..., schiaveg, Apr-24-02, (10) RE: High background in Amino-allyl ..., Tineke, May-07-03, (11) RE: High background in Amino-allyl ..., carriehung, Jun-26-03, (13) RE: High background in Amino-allyl ..., yuhua, Jul-24-03, (14) RE: High background in Amino-allyl ..., raclaus, Jun-20-03, (12)

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Chantal Aug-23-01, 06:20 PM (PDT) 1. "RE: High background in Amino-allyl labelling " Hi Masako, I've been working with the aminoallyl protocol for a while, and my background is very low. I can email you my protocol, if you give me an address. Chantal   Remove | Alert Edit | Reply | Reply With Quote | Top

masako Aug-26-01, 08:59 PM (PDT) 2. "RE: High background in Amino-allyl labelling " Thank you so much, that will be wonderful. I am kind of suspecting that it may be slide processing but I will be grateful if you could email me your protocol. my email address is masako@anatomy.usyd.edu.au Thank you again Masako   Remove | Alert Edit | Reply | Reply With Quote | Top

zhengd Mar-27-02, 08:23 AM (PDT) 3. "RE: High background in Amino-allyl labelling " Dear Chantal: I am very interested in your protocol since I also got high background. Could you please email me a copy of your protocol. My email address is DXZ082@bham.ac.uk By the way, have you also met problem about inconsistent labelling with cy3/cy5? My cy5 signal is always much lower than cy3 signal   Remove | Alert Edit | Reply | Reply With Quote | Top

cyj Apr-04-02, 02:06 PM (PDT) 4. "RE: High background in Amino-allyl labelling " I also got the similiar problem. Could you please send me a copy of your protocol? My email address is cui1989@yahoo.com Thank you very much.   Remove | Alert Edit | Reply | Reply With Quote | Top

marchini Apr-13-02, 02:27 PM (PDT) 5. "RE: High background in Amino-allyl labelling " Hello Chantal This is sergio and I red your mail to Masako and since i 'm involvrd in indirect labelling i would be interest in reading your protocol..... could you please mail me a copy please All the best sergio   Remove | Alert Edit | Reply | Reply With Quote | Top

mojgana Apr-16-02, 09:45 AM (PDT) 6. "RE: High background in Amino-allyl labelling " Hi Chantal, could you please e-mail me your aminoallyl protocol . Thank you Mozhgan   Remove | Alert Edit | Reply | Reply With Quote | Top

Chantal Apr-18-02, 03:34 PM (PDT) 7. "RE: High background in Amino-allyl labelling " Hello Sergio and Mozhgan, I'm willing to send my protocol (that is based on Joe deRisi's protocol), but I need an email address! Chantal   Remove | Alert Edit | Reply | Reply With Quote | Top

marchini Apr-21-02, 11:07 AM (PDT) 8. "RE: High background in Amino-allyl labelling " dear Chantal Thanks a lot for your replay and forgive me for my delay in answering to you. My email address is marchini@marionegri.it In those days I'm doing different arrays from Clontech (Atlas Human Arrays 3.8 K genes) based on indirect labelling. They are working very well but in my opinion washing should be optimised to reduce background..... all the best Sincerely yours Sergio   Remove | Alert Edit | Reply | Reply With Quote | Top

alli Apr-23-02, 01:02 AM (PDT) 9. "RE: High background in Amino-allyl labelling " Hi Chanttal, Can you also email me a copy of your protocal (stackaj@hotmail.com). I am having trouble with a low signal. Thanks, Allison   Remove | Alert Edit | Reply | Reply With Quote | Top

schiaveg Apr-24-02, 07:25 AM (PDT) 10. "RE: High background in Amino-allyl labelling " Hi Chantal, I would appreciate receiving your protocol. E-mail: schiaveg@usp.br Thanks a lot. Silvana   Remove | Alert Edit | Reply | Reply With Quote | Top

Tineke May-07-03, 03:15 AM (PDT) 11. "RE: High background in Amino-allyl labelling " Hi Chantal, I wonder whether the background problems relate to the type of coating on the glass slides, so do you use poly-L-lysine-coated glass slides for your amino-allyl labeled samples? Tineke   Remove | Alert Edit | Reply | Reply With Quote | Top

carriehung Jun-26-03, 06:32 AM (PDT) 13. "RE: High background in Amino-allyl labelling " Hi Chantal, I am working on amino-allyl labelling as well. Do you mind send me your protocol? Thanks a lot! Carrie   Remove | Alert Edit | Reply | Reply With Quote | Top

yuhua Jul-24-03, 03:40 PM (PDT) 14. "RE: High background in Amino-allyl labelling " Hi, Dear Chantal I am working on indirect labelling and interested in your protocol. Could you please send me a copy? my email address is zou@salk.edu. Thanks! Best regards, yuhua   Remove | Alert Edit | Reply | Reply With Quote | Top

raclaus Jun-20-03, 06:08 AM (PDT) 12. "RE: High background in Amino-allyl labelling " Dear Chantal, My name is R. Claus, I'm also working on microarray hybridizations and I've got recently the same problem. Could you, please, send my your modified/optimized protocol too. I would very appreciate it. My e-mail address: ralf.claus@med.uni-jena.de Many thanks in advance, Sincerely Ralf Claus   Remove | Alert Edit | Reply | Reply With Quote | Top

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