| Description |
| SABiosciences' GEAhyb Hybridization
Solution is an optimized hybridization buffer prepared for use with all
cDNA Q and S Series GEArrays® and Oligo GEArrays® The GEAhyb
Hybridization Solution is especially suited to our nylon-based microarray
technology. GEAhyb Hybridization Solution is used to hybridize labeled DNA
or RNA in microarray and dot-blot experiments. For the cDNA Q and S Series
GEArrays, it is also used with sheared salmon sperm DNA (available
separately as catalog number GA-007) in the preparation of GEAprehyb
blocking solution. The GEAhyb Hybridization Solution provides enough
material to process 12 GEArray membranes. Instructions for the use of
GEAhyb Hybridization Solution are included in your GEArray User Manual. A
brief protocol for experienced array users is also provided below.
Need more information about GEArray products? please visit Microarray Home or send Email to Technical Support |
| Kit Contents /
Packing List / Storage Conditions |
Please check the kit components immediately after you receive this
package. SABiosciences will replace any missing items if reported within two
(2) business days of receipt.
Contents: One bottle GEAhyb Hybridization Solution
Storage Conditions:
GEAhyb Hybridization Solution is shipped at ambient temperature. Store at room temperature.
Shelf Life: All reagents are stable for 2 years after receipt of the
kit if stored at the recommended temperature.
NOTE: Some components of the solution will precipitate at lower
temperatures. This will not affect performance of the hybridization solution
providing that all buffer components are completely dissolved before you use
the solution. To ensure complete dissolution of buffer components take
special care in following the GEArray User Manual instructions (involves
pre-warming the solution to 60 ºC for 30 minutes)
|
| Brief Protocol: For
Experienced Users |
Pre-Hybridization
- Pre-wet the array membrane with 5 ml deionized water for 5 min in
the provided hybridization tube. Screw on the cap hand-tight and allow
tube to sit inverted while preparing the GEAprehyb solution.
- Warm the GEAhyb Hybridization Solution to 60 ºC and invert the
bottle several times to allow complete dissolution of the buffer
components.
- For cDNA GEArrays only, heat sheared salmon sperm DNA at 100 ºC for
5 min. Chill on ice immediately. Prepare GEAprehyb: Add the
heat-denatured salmon sperm DNA to the pre-warmed GEAhyb Hybridization
Solution to a 100 µg/ml final concentration. You will need 3 ml of
GEAprehyb for each array. Keep the GEAprehyb solution at 60 ºC until
needed.
- For Oligo GEArrays only, use 2 ml pre-warmed (60 ºC) GEAhyb
Hybridization Solution directly as the pre-hybridization (GEAprehyb)
solution instead.
- Discard the deionized water from the hybridization tube. Add 2 ml of
the appropriate GEAprehyb solution, and vortex the tube gently for a
few seconds. Be sure the cap of the tube is screwed on hand-tight.
- Place the tube inside your hybridization cylinder.
- Pre-hybridize in a hybridization oven at 60 ºC for 1 to 2 hours
with continuous agitation at 5 to 10 rpm.
Hybridization:
- Prepare GEAhyb: Add target or probe (labeled cDNA or cRNA for cDNA
or Oligo GEArray, respectively) to 0.75 ml of pre-warmed GEAprehyb
for the cDNA GEArray or 0.75 ml of pre-warmed GEAhyb Hybridization
Solution for the Oligo GEArray. Mix well, and keep the GEAhyb at 60
ºC.
- Discard the GEAprehyb from the hybridization tube.
- Add the GEAhyb with probe/target to the hybridization tube.
- Hybridize overnight at 60 ºC with continuous agitation at 5 to 10
rpm.
- Continue to the wash and detection steps of the microarray
experiment.
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