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Old 08-06-2009, 06:40 PM   #1
AKroxy
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Default suppression subtractive hybridization and 454

Has anyone done suppression subtractive hybridization PCR (SSH) on cDNA prior to 454 sequencing? Any tips, protocols, etc would be greatly appreciated.
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Old 08-31-2009, 04:28 AM   #2
pmiguel
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Quote:
Originally Posted by AKroxy View Post
Has anyone done suppression subtractive hybridization PCR (SSH) on cDNA prior to 454 sequencing? Any tips, protocols, etc would be greatly appreciated.
Probably not what you are looking for, but the following uses an Evrogen kit to normalize cDNA libraries prior to 454 sequencing:

http://www.pubmedcentral.nih.gov/art...?artid=2689275

Evrogen also sells SSH reagents, so that could be a place to look.

Phillip
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Old 01-14-2011, 01:47 AM   #3
verdemalaquita
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Default ssh + 454

Hello,
Do you know if are there some information about supression substrative hubridization using clontech kit prior to pyrosequencing 454???
Thanks in advance!!!
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Old 01-14-2011, 11:06 AM   #4
AKroxy
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I tried tweaking that protocol to get rid of homopolymers so we could use the samples in 454 sequencing, but I failed. I would have needed to spend a lot more time on it to get it to work.
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