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Genome walking - (Sep/02/2004 )

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QUOTE (Sequencer @ Mar 22 2005, 12:47 AM)
[i hope you could elaborate on the TSPs... I am also interested of making use of seegene... how much did it cost? thanks!


TSPs are just primers you will use to amplify unknow DNA sequence with a link primer in genome walking. You can design it by yourself and order it from any company.

-pcrman-

QUOTE (pcrman @ Mar 22 2005, 07:41 PM)
TSPs are just primers you will use to amplify unknow DNA sequence with a link primer in genome walking. You can design it by yourself and order it from any company.


i aslo sent this messeage to without00.

i am using seegene kit, but i've got nonspecific sequence. it is amplified by ACP primers. so i am wondering how too design an effective TSP primer.

according to the mannual, ACP primer-binding sites(5-XGGTC-3) upstream of TSP1 or in/between TSP primers should be avoided. why is this a big deal? there must be a limit of distance for the distance. i mean we should only consider a certain distance upstream, like within 1kb. am i right?

how about those binding sites downstream of the TSP primers? doesn't matter?

how long fragment can you get using this kit? can we use long PCR system instead.

hope you can share your expereince with us.

thanks a lot.

-seqman-

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