10. Washing unbound probe from glass array (NON-Automated version).

 

Remove chamber cover from probed array over garbage can as hybridization solution will drip.

Carefully remove any remaining adhesive material from the array with a razor blade.

Places arrays in Wheaton glass slide holder.

Wash in pre-warmed 2X SSC, 0.2% SDS washing solution at 65oC for 5 minutes.

(Every minute lift the tray up and down 2-3x to aid in washing)

Wash the array in 2X SSC, 0.2% SDS washing solution at RT for 2 minutes.

Wash the array in 0.2X SSC, 0.2% SDS washing solution at RT for 2 minutes.

Wash the array in 0.2X SSC, 0.2% SDS washing solution at RT for 2 minutes.

Air Dry the array.

 

11. Hybridization of Dendrimers (Cy3 and Cy5) to Array (NON-Automated version).

 

Place circular filter papers (about 3-4) into both reservoirs of pre-warmed incubation chamber.

Add 40-50 ul of 2X SSC onto each filter paper stack.

Place the air-dried array into the chamber. (DNA side up)

Add 2 ul of 65oC dendrimer mixture into 13 ul of 65oC 3DNA hybridization buffer and mix well.

(These values are for a 25 mm x 25 mm cover slip)

(Double the volumes for a 40 mm x 25 mm cover)

Place the 15 ul hybridization solution and dendrimers on the center of the array.

(Between the scratches made with the diamond pencil)

Carefully place the ethanol wiped and dried glass cover slip (Corning) on the drop of solution.

If necessary, remove bubbles by gentle tapping on glass with a pipette tip.

Assemble the incubation chamber cover on the base. (MAKE SURE GASKET IS SEATED)

Apply a binder clip at each end, and fold the metal hinges toward the middle.

Place the chamber and array at 65oC for 6 hours.

 

12. Washing unbound dendrimer from glass array(NON-Automated version).

 

Remove incubation chamber from 65oC incubator.

Carefully remove binder clips and cover from incubation chamber.

Places arrays in Wheaton glass slide holder.

Wash in pre-warmed 2X SSC, 0.2% SDS washing solution at 65oC for 10 minutes AFTER COVER

SLIP FALLS OFF!

(Every minute lift the tray up and down 2-3x to aid in washing)

(Make sure that the cover slip falls off, do not help in any other way)

Wash the array in 2X SSC, 0.2% SDS washing solution at RT for 2 minutes.

Wash the array in 0.2X SSC, 0.2% SDS washing solution at RT for 2 minutes.

Wash the array in 0.2X SSC, 0.2% SDS washing solution at RT for 2 minutes.

Air Dry the array.

Wipe the back of the array with a Kimwipe (70 % ETOH treated).

Scan the array.

 

 

 

 

                                                                                                                               

 
 

Microarray Protocols

 

Basic knowledge of microarray.

 

Basic knowledge of microarray 2.

 

Introduction to Microarray.

 

A Mediabook of Microarrays

 

Protein chips 

 

MicroArray Procedure

(Modified by Ryoji Kojima, Takumi Yoshida and Jeffrey Randall, 8.20.2000)

 

1. Total RNA Isolation from cultured cells.

 

2. DNase Treatment of Total RNA

 

3. Making the single strand cDNA probe.

 

4. Automated Slide Processor (ASP) Version for hybridization.

 

5. Washing microarrays in ASP.

 

6.Processing of Array slide

 

7. Pre-hybridization of the processed slides (NON-Automated version).

 

8. Hybridization of Cy3 + Cy5 probe to glass array (NON-Automated version).

 

9. Preparation of Dendrimer Cy3 and Cy5.

 

10. Washing unbound probe from glass array (NON-Automated version).

 

11. Hybridization of Dendrimers (Cy3 and Cy5) to Array (NON-Automated version).

 

12.Washing unbound dendrimer from glass array(NON-Automated version).

 

Microarray Dababases

 

Troubleshooting

 

Other Microarray Protocols (1)

 

Other Microarray Protocols (2)

 

Other Microarray Protocols (3)

Web Guider

Ch 1.General Lab Techniques

Ch 2.Molecular Separation

Ch 3.DNA and RNA

Ch 4.Genetics

Ch 5.PCR Serials

Ch 6.Protein

Ch 7.DNA Protein Interactions

Ch 8.Immunohistoch / immunology

Ch 9.Cellular Biology

Ch 10.GC/MS, NMR and Proteomics

Ch 11.Animal Experiments

Ch 12.Worm: C. Elegans

Ch 13.HPLC and TLC

Ch 14.Buffers formats in Lab.

Ch 15.Other Resources

Free eBooks at Library Online

Cinema Online,Free Movies-(1)

Progresses in Life Science

Free eBooks in biomedicine

Pathway databases

Biological Educational Resources

Textbooks and Lab Manuals

Lab-Manual.Com
Home || Biosocial || Videos || Recipes || Resource