# Sequencing

**URL:** https://forum.mothur.org/t/sequencing/1344
**Category:** Theory behind mothur
**Created:** [May 14, 2013, 4:03pm UTC](https://forum.mothur.org/t/sequencing/1344 "2013-05-14T16:03:35Z")
**Posts on this page:** 3
**Page:** 1

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### Author: ![nminalt](https://avatars.discourse-cdn.com/v4/letter/n/90db22/32.png) [@nminalt](https://forum.mothur.org/u/nminalt)
#### Post date: [May 14, 2013, 4:03pm UTC](https://forum.mothur.org/t/sequencing/1344/1 "2013-05-14T16:03:35Z")

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When we get our sequences pyrosequenced using 454 we have them do it off the 28F primer. Are our sequences then running 5’-3’ or 3’-5’?

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### Author: ![pschloss](https://yyz2.discourse-cdn.com/flex036/user_avatar/forum.mothur.org/pschloss/32/4_2.png) [@pschloss](https://forum.mothur.org/u/pschloss)
#### Post date: [May 14, 2013, 5:12pm UTC](https://forum.mothur.org/t/sequencing/1344/2 "2013-05-14T17:12:44Z")

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5-\>3 = so you do not need to flip them

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### Author: ![nminalt](https://avatars.discourse-cdn.com/v4/letter/n/90db22/32.png) [@nminalt](https://forum.mothur.org/u/nminalt)
#### Post date: [May 14, 2013, 6:45pm UTC](https://forum.mothur.org/t/sequencing/1344/3 "2013-05-14T18:45:06Z")

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Thank you!