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AW: [Automated-sequencing] geneScan 500-Liz or geneScan 500-Rox

Axel Kuenzler via autoseq%40net.bio.net (by info from advancedbiolabservice.com)
Tue Oct 14 03:20:39 EST 2014


Hello Myriam,

Please check the official available combinations of dyes and the
corresponding Matrix Standard kits for ABI310/377 or ABI multi-capillary
sequencers:
http://www.advancedbiolabservice.com/Temp/DyeSets_for_Genotyping.pdf

Normally the last (= 4th or 5th) dye is reserved for the size standard
unless you define something else. The other (3 or 4) dyes can be used for
multiplex/multicolor fragment analysis. LIZ labelled primers are unusual.

Combinations with your dyes would be:
DS-33/G5 = 6FAM, VIC, NED and PET combined with LIZ size standard DS-31/D =
6FAM, VIC and NED combined with ROX size standard

ROX and PET are both "red" colors and can not be analysed at the same time.
If you run (G5 dye set) fragments with 6FAM, VIC, NED and LIZ and analyze
them with ROX size standard you will get little cross talk of the ROX
sizestandard into the yellow NED channel (and possibly also little green)
which is depending on your type of analysis possibly acceptable. If you are
using ABI 310, you can create your own matrix matching exactly with 6FAM,
VIC, NED, ROX, LIZ (by running some ROX fragments instead of PET fragments).

Next time do not order the expensive original ABI colors, but alternative
cheaper 5-dye color dyes like: 5/6FAM, Yakima Yellow, ATTO 550, ATTO 565 and
ATTO 633


Good luck,
AXEL

-----Ursprüngliche Nachricht-----
Von: autoseq-bounces from oat.bio.indiana.edu
[mailto:autoseq-bounces from oat.bio.indiana.edu] Im Auftrag von
myriamharrabi.mh from gmail.com
Gesendet: Montag, 13. Oktober 2014 11:51
An: bionet-genome-autosequencing from moderators.isc.org
Betreff: [Automated-sequencing] geneScan 500-Liz or geneScan 500-Rox

Hi all,
i used to work with geneScan 500-Liz and 6FAM and Ned dyes.
here i am dealing with genotyping. i commanded dyed primers with 5
differents dyes: 6FAM, VIC, NED PET ans Liz. at the same time, actually i
can just offord GeneScan 500-Rox.
Can i work with GeneScan 500-Rox using the dyes cites below?
All the best
MH

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