Biomedical Engineering Reference
In-Depth Information
(RT, 10 sec), Universal Pre-Hybridization Solution (42
◦
C, 15 minutes),
Wash Solution 2 (RT, 1 minute), Wash Solution 3 (RT, 30 sec)
dry slides by low speed spin (1000 rpm) at RT, 1 minute
4.
Hybridization
•
•
dissolve labelled cDNA in 60
µ
l Universal Hybridization buffer
denature the target mixture at 95
◦
C for 3 minutes, then spin for 20 sec
at RT
•
•
place onto the array, cover with 24
×
60 mm cover-slip (avoid bubbles!)
incubate in a high humidity hybridization chamber at 42
◦
C, 14-20
hours
5.
Post hybridization processing
•
•
immerse slides in Wash Solution 1 (2 minutes), remove coverslip, then
incubate for 5 minutes, all at 42
◦
C
transfer slides successively to Wash Solution 1 (5 minutes, 42
◦
C), Wash
Solution 2 (10 minutes, RT), wash solution 3 (2 minutes, RT), wash
solution 3 (2 minutes, RT)
•
•
dry slides by low speed spin (1000 rpm) for 1 minute at RT; store in
the dark
References
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Sequenc-
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.Adv
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AD. 1989.
An oligonucleotide hybridization approach to DNA sequencing
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.
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