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-   -   Pe150+8(+8?)+150 (http://seqanswers.com/forums/showthread.php?t=93142)

mdegraaf 02-25-2020 12:11 PM

Pe150+8(+8?)+150
 
Hi,
We have prepared a bunch of libraries with the Nextera Flex kit, with the use of the index plate (Nextera™ DNA CD Indexes (96 Indexes, 96 Samples). Now we are sending pooled libraries off to be sequences and we have to select if we want PE150+8+150 or PE150+8+8+150. Unfortunately there is no further information on what this means. Does anyone know? I would think the double 8 because of dual indexing but would appreciate any input

GSviral 02-25-2020 10:58 PM

Hello mdegraaf,

You are correct with your assumption. The PE 8 + 8 option refers to a dual indexing strategy while the PE 8 option will just be a single indexing approach. However, if you are still concerned I would advise getting in contact with the sequencing provider and they can put your mind at ease fairly quickly.

Hope this helps!

mdegraaf 02-26-2020 01:20 PM

Thank you for your answer. I did contact the provider but haven't heard back from them yet.

kmcarr 02-27-2020 08:01 AM

mdegraaf,

You didn't say how many libraries you pooled. If it is more than 12 then it is inevitable that you have more than one library sharing a common i7 index sequence. i7 is the first of the two index sequenced or the ONLY index sequenced if you choose PE150+8+150. You can not use this format if any i7 index is repeated in your pool, you must then use PE150+8+8+150. Honestly I'd always choose the dual index sequencing format; what would be the downside to it?


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