Miseq compatible in house library preparation method

vl80 · 12-10-2012, 09:28 PM

Dear all,

We just go a quotation for our Miseq runs and the cost of the TruSeq kit is way too high for regular use as we intend to use this in long term
1. Are there any published methods of in house library prepation that is Miseq compatible?
2. Are Miseq adaptors the same as Hiseq? I have found a document with Hiseq adaptor sequences but not Miseq
3. What are the cheaper alternative library preparation kits availabe?

We plan to sequence PCR amplicons than long fragments so would not be needing a shearing step.
Any suggestions are welcome or rather we are desperate for ideas!
Thanks a lot

pmiguel · 12-11-2012, 03:28 AM

Quote:

Originally Posted by vl80

Dear all,

We just go a quotation for our Miseq runs and the cost of the TruSeq kit is way too high for regular use as we intend to use this in long term
1. Are there any published methods of in house library prepation that is Miseq compatible?
2. Are Miseq adaptors the same as Hiseq? I have found a document with Hiseq adaptor sequences but not Miseq

Yes, they are the same.

Quote:

Originally Posted by vl80

3. What are the cheaper alternative library preparation kits availabe?

We plan to sequence PCR amplicons than long fragments so would not be needing a shearing step.

You will not need library construction then. Just use a 'fusion primer' or 'step out PCR' method.

vl80 · 12-11-2012, 04:15 AM

Thank you very much for the reply.
One more thing, my PCR is actually a multiplex with 23 primer pairs, already optimised. Therefore, adding barcodes and adaptors to the primary PCR primers would be difficult.
Experience sharing with step out PCR instead of library prep is very welcome.

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12-10-2012, 09:28 PM	#1
vl80 Member Location: Asia Join Date: Nov 2012 Posts: 34	Miseq compatible in house library preparation method Dear all, We just go a quotation for our Miseq runs and the cost of the TruSeq kit is way too high for regular use as we intend to use this in long term 1. Are there any published methods of in house library prepation that is Miseq compatible? 2. Are Miseq adaptors the same as Hiseq? I have found a document with Hiseq adaptor sequences but not Miseq 3. What are the cheaper alternative library preparation kits availabe? We plan to sequence PCR amplicons than long fragments so would not be needing a shearing step. Any suggestions are welcome or rather we are desperate for ideas! Thanks a lot

12-11-2012, 04:15 AM	#3
vl80 Member Location: Asia Join Date: Nov 2012 Posts: 34	Thank you very much for the reply. One more thing, my PCR is actually a multiplex with 23 primer pairs, already optimised. Therefore, adding barcodes and adaptors to the primary PCR primers would be difficult. Experience sharing with step out PCR instead of library prep is very welcome. Last edited by vl80; 12-11-2012 at 12:13 PM.