[pmiguel]

Does anyone know of alternatives to Life Technologies for this reagent? Since their acquisition of ABI, our cost has nearly doubled, and the costs for other reagents have increased as well.

Thanks!

Barry

Ethanol precipitation. But you need to seriously cut back on your Big Dye because EtOH isn't as effective at removing persisting reactants (dye blobs). Also, we generally use less than 2 volumes of ethanol to reduce the final salt concentration and reduce the predominance of short products. May not be as critical if you load in formamide -- we always do water loads for the higher signal strength. Oh, you also need a centrifuge, so it may not be ideal for your circumstances.

--
Phillip

[Dynamac]

Thanks, all.

[Vinz]

BD V1.1 is supposed to be better for the short products than BD V3.1
We use POP6 on one of our 3730XL for reading early. Using BD V1.1 and Pop6 reading from base 20 onwards is no problem. If you correct it manually, you could read from base 1 often. Caveats: Pop6 is only sold in very small aliquots and with a huge price tag.
If you are interested I could share the settings.

[Dynamac]

I was under the impression that Pop 6 would not work on the 3730, but I'm glad to hear otherwise. If you could share the settings with me that would be great!

You can send a private message if you'd like, or just post them here.

Thanks, Vinz.

[Vinz]

We got a patch from ABI to allow for POP6. If I recall correctly the main thing is a new POP6 mobility file to be selected in the "analysis protocol". But I think this is not required.
Oven temperature is set to 55°C
PreRun and Run Voltage at 15kV (13kV gives a little better resolution, but takes longer)
160ms readoutime (first and second)

I would recommend a new spectral calibration after changing the POP.

[pmiguel]

BD V1.1 is supposed to be better for the short products than BD V3.1
We use POP6 on one of our 3730XL for reading early. Using BD V1.1 and Pop6 reading from base 20 onwards is no problem. If you correct it manually, you could read from base 1 often. Caveats: Pop6 is only sold in very small aliquots and with a huge price tag.

Ah, I remember when we bought it in big (100 ml?) bottles for the 3700 -- $300 each. What a shame for Applied Biosystems that manufacturing POP-6 is so expensive that they have been forced to increase their prices by nearly three orders of magnitude. I am sure they are weeping bitter tears over that.

If you are interested I could share the settings.

Please post them here.

--
Phillip

[Vinz]

There is an alternative source for the POP6:

http://www.mclab.com/NanoPOP-TM-Polymers.html

However, so far we did not take the effort and risk of testing if those perform similar...
If you test those we would be interested in your experience.

[MrGuy]

Yep, I heard about those, too. I haven't heard anything bad about them. The company apparently regenerate arrays after they "go bad".