Download PerfectHyb hybridization solution

www.toyobo.co.jp/e/bio
(4) Probe concentration
The following table lists recommended probe concentrations. Higher-concentrated
probes tend to generate background signals, in particular with analyses employing
non-radioactive probes.
Label
RI
Non-RI
Probe concentration
2.5–5 pmoles/ml
0.5–1 pmoles/ml
(5) Protocol
(a) Prehybridize 10 x 10 cm membranes in a minimum total volume of 5 ml of
PerfectHyb hybridization solution at optimal hybridization temperature for 20
minutes.
(b) Add the labeled probe to 5 ml of prewarmed, fresh PerfectHyb hybridization
solution*.
*Assure that probe is thoroughly mixed with fresh hybridization solution.
(c) Replace pre-hybridization solution with fresh hybridization solution containing
the labeled probe.
(e) Incubate at optimal hybridization temperature for 1–2 hours.
(f) Wash membrane two times in pre-warmed Wash solution 1 at the optimal
temperature for 5 minutes.
(g) Wash membrane two times in pre-warmed Wash solution 2 at the optimal
temperature for 10 minutes.
(h) [Radioactive probe]
Transfer membrane with forceps to filter paper. Then, follow with exposure to
X-ray film at -70°C.
[Non-radioactive probe]
Perform detection according to the instruction manual.
JAPAN
TOYOBO CO., LTD.
Tel(81)-6-6348-3888
www.toyobo.co.jp/e/bio
[email protected]
CHINA
TOYOBO Bio-Technology, CO., LTD.
Tel(86)-21-58794900.4140
FOR RESEARCH USE ONLY.
6
NOT FOR HUMAN OR DIAGNOSTIC USE.