Reusing Microarray Chip - (May/28/2014 )

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 95^oC sequentiially.

Thanks..

-Inbox-

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.

-mdfenko-

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

http://www.kellogg.umich.edu/bios/downloads/kahana/SouthernBlotting.pdf

What could be reason for less intense and sensitive second hybridization?

-Inbox-

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.

-mdfenko-