Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

Reusing Microarray Chip


  • Please log in to reply
3 replies to this topic

#1 Inbox

Inbox

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 331 posts
21
Excellent

Posted 28 May 2014 - 03:12 AM

Hi,

   I want to  reuse microarrray chip again for next hybridization experiment. Please let me know if I can and what coud be protocol?

 

Right now I am thinking just having heating wash buffer to 95 oC. Keeping five flask and then transferring chip in five flask of 95oC sequentiially. 

 

Thanks..


Edited by Inbox, 28 May 2014 - 03:13 AM.


#2 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 3,248 posts
198
Excellent

Posted 28 May 2014 - 03:38 AM

which brand chip?

 

if agilent, did you use stabilizer? if so then you may not be able to properly strip for reuse (just a guess).

 

if not then you should be able to strip the same way you would strip any hybridization (eg southern blot). the only caveat (as was told to me by an employee of agilent, so a grain of salt should be taken with the advice) is that the second hybridization result may be less intense (and, thereby, less sensitive) than the first.


talent does what it can
genius does what it must
i do what i get paid to do

#3 Inbox

Inbox

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 331 posts
21
Excellent

Posted 28 May 2014 - 08:34 AM

Agilent chip.

I did not used stabilizer.

 

Do you mean, I can follow following protocol which is used for southern blot, 

Stripping solution

1. Wash array in water for 2-5'. 
2. Strip with 500 ml stripping solution (0.2N NaOH 0.1% SDS - made fresh) at 37 deg C for 30'. 
3. Wash with 2x SSC at RT/1'. Keep in SSC until ready to use. 
 
20x SSC 
 3M NaCl 
 0.3M Na-citrate; pH7.0
 
 
What could be reason for less intense and sensitive second hybridization?

Edited by Inbox, 28 May 2014 - 10:03 AM.


#4 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 3,248 posts
198
Excellent

Posted 29 May 2014 - 03:36 AM

you shouldn't need to keep in ssc. the arrays are stored dry, so you should wash with water and dry (you can use the ssc first to ensure that the stripping solution is neutralized and removed).

 

i wasn't given an explanation but i would guess that the reason why the result may be less intense could be due to incomplete stripping of the first sample and possibly (but not really likely, in my mind) damage to the probes.


Edited by mdfenko, 29 May 2014 - 03:37 AM.

talent does what it can
genius does what it must
i do what i get paid to do




Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.