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Forum: Illumina/Solexa 10-19-2017, 05:58 AM
Replies: 17
Views: 2,122
Posted By suefo
Hi jteeee2, I just wanted to check back in with...

Hi jteeee2,
I just wanted to check back in with you and say thank you. We did some testing in the lab and both the primers / Taq you recommended appeared to work well. We submitted that to our...
Forum: 454 Pyrosequencing 09-09-2013, 10:52 AM
Replies: 3
Views: 2,190
Posted By suefo
We have a MiSeq... but we need to read across...

We have a MiSeq... but we need to read across larger regions.
Forum: 454 Pyrosequencing 09-09-2013, 08:32 AM
Replies: 3
Views: 2,190
Posted By suefo
FLX+ and amplicons

Good afternoon all. I understand there are people out there now using the FLX+ with amplicons (which is now fully supported by 454 / Roche). We are contemplating an upgrade from FLX to FLX+ but...
Forum: 454 Pyrosequencing 03-28-2013, 10:06 AM
Replies: 5
Views: 1,627
Posted By suefo
By the way, I just checked Fishers catalog and...

By the way, I just checked Fishers catalog and they have them in stock (the new tips). Sue
Forum: 454 Pyrosequencing 03-28-2013, 10:04 AM
Replies: 5
Views: 1,627
Posted By suefo
Afternoon! These worked for us (the new catalog...

Afternoon!
These worked for us (the new catalog number) - hopefully those aren't the ones discontinued as well. We use them for both the FLX as well as the JR.
Forum: 454 Pyrosequencing 02-28-2013, 11:10 AM
Replies: 0
Views: 1,421
Posted By suefo
Updates on FLX+ and amplicons

Hello all,
I've been away from the forum for some time running libraries like crazy. I was just wondering if anybody has updated information on support from Roche for amplicons using the FLX+...
Forum: 454 Pyrosequencing 10-23-2012, 11:51 AM
Replies: 5
Views: 1,627
Posted By suefo
Combitips

So last year, after many months of struggling, it came down to our high % enrichment being caused for the Matrix pipettor we were using. When we switched over to an Eppendorf Repeater Pipettor,...
Forum: 454 Pyrosequencing 09-25-2012, 04:31 AM
Replies: 5
Views: 1,575
Posted By suefo
FLX+ and amplicons

Hi all, I am hoping for a lot of feedback. We don't have a sales rep or application scientist working our region, so I could really use updates on how well the FLX+ upgrades are working with...
Forum: 454 Pyrosequencing 03-21-2012, 10:11 AM
Replies: 18
Views: 7,800
Posted By suefo
Christina85... I am ABSOLUTELY sure it was the...

Christina85... I am ABSOLUTELY sure it was the pipettor. I have photos and data to support this is a side-by-side experiment we did. We checked everything before hand as well... pcr plate, thermal...
Forum: 454 Pyrosequencing 11-18-2011, 10:25 AM
Replies: 18
Views: 7,800
Posted By suefo
Anthony, The Matrix was VERY slow and yes, on...

Anthony,
The Matrix was VERY slow and yes, on the final pickup, we overshot the volume on the draw. The repeator is much much faster - we don't try to slow it down. We just shoot into the wells...
Forum: 454 Pyrosequencing 10-14-2011, 10:56 AM
Replies: 2
Views: 3,838
Posted By suefo
Innovative Solutions vs AMPure

Today we were visited by folks from Innovative Solution who were promoting their PCRClean and DNA SizeSelect purification kits. They claim that side-by-side with AMPure, we would get far better...
Forum: 454 Pyrosequencing 09-26-2011, 04:45 AM
Replies: 18
Views: 7,800
Posted By suefo
We're using 1 mL combi-tips for SVs, 2.5 mL...

We're using 1 mL combi-tips for SVs, 2.5 mL combi's for MVs and 5 mL combi's for LVs. And so far the diameter of the barrel seems to fit.
Forum: 454 Pyrosequencing 09-26-2011, 04:09 AM
Replies: 18
Views: 7,800
Posted By suefo
OK.. I know this is going to sound silly but it's...

OK.. I know this is going to sound silly but it's true. Originally we were using a Matrix pipette (set at the slowest possible speed) to transfer our emPCRs to the plate and we got terrible...
Forum: 454 Pyrosequencing 09-22-2011, 05:26 AM
Replies: 18
Views: 7,800
Posted By suefo
We run many GS Junior emPCR Lib-L kits. When...

We run many GS Junior emPCR Lib-L kits. When determining % initial bead recovery we use the 10e6 number. However, we adjust our %enrichment based on the recovered beads and not the original 10e6. ...
Forum: 454 Pyrosequencing 09-22-2011, 05:21 AM
Replies: 0
Views: 1,845
Posted By suefo
HP Z400 computer issues?

Well the running joke in our lab was everything on our GS Junior had been replaced except for the camera - when I pointed out that the computer was still original, too. Then of course, the next run,...
Forum: 454 Pyrosequencing 08-23-2011, 11:41 AM
Replies: 0
Views: 1,244
Posted By suefo
Eppendorf Repeater Plus Pipettor vs Matrix Programmable

So we just completed a side-by-side experiment where the only difference was how the emulsions were delivered to the PCR plate... one with a repeater pipettor and the other with a matrix programmable...
Forum: 454 Pyrosequencing 08-19-2011, 03:47 AM
Replies: 17
Views: 3,437
Posted By suefo
Yes, Junior kits. I'm stumped as these tend to...

Yes, Junior kits. I'm stumped as these tend to be the most consistent in our hands.
Forum: 454 Pyrosequencing 08-19-2011, 03:44 AM
Replies: 6
Views: 3,093
Posted By suefo
Our EBox went last week from the machine we got...

Our EBox went last week from the machine we got in March. Now the valve block has gone.
Forum: 454 Pyrosequencing 08-18-2011, 11:01 AM
Replies: 17
Views: 3,437
Posted By suefo
Are you using LVs, MVs, SVs or JR kits? Our JR...

Are you using LVs, MVs, SVs or JR kits? Our JR kits tend to be consistent. But we see variation on the others - especially the MVs.
Forum: 454 Pyrosequencing 08-18-2011, 06:21 AM
Replies: 6
Views: 3,093
Posted By suefo
runAnalysisFilter on GS Junior computer

This should only take 2 steps (see attached jpg)/
Forum: 454 Pyrosequencing 08-12-2011, 12:09 PM
Replies: 9
Views: 3,380
Posted By suefo
I will send them to you. You should be able to...

I will send them to you. You should be able to see them yourself. They should be under DOCUMENTS on the my454.com/my454 window. The last item listed in that drop down menu is Technical Bulletins. ...
Forum: 454 Pyrosequencing 08-12-2011, 04:08 AM
Replies: 9
Views: 3,380
Posted By suefo
Take a look at TCB 2011-001. I spoke with 454...

Take a look at TCB 2011-001. I spoke with 454 this week about this issue and they recomended this bulletin. Even though it reads for Lib-A, 454 felt it would also work with Lib-L. BUT you will...
Forum: 454 Pyrosequencing 08-08-2011, 03:54 AM
Replies: 7
Views: 2,677
Posted By suefo
Ooops... I stand corrected on our length. I just...

Ooops... I stand corrected on our length. I just re-checked the agilent data and it appears the DNA being submitted for testing is 650 bp (that's including the PCR adaptors, barcodes and keys).
Forum: 454 Pyrosequencing 08-08-2011, 03:48 AM
Replies: 7
Views: 2,677
Posted By suefo
Thanks again RCJK. Our %Short is around 60%...

Thanks again RCJK.
Our %Short is around 60% (%Dot+Mix is at 15%).
We do perform an initial AMPure, run the products on the Agilent, pool DNA, re-purify a second time using AMPure and then run a...
Forum: 454 Pyrosequencing 08-07-2011, 04:51 PM
Replies: 7
Views: 2,677
Posted By suefo
RCJK... thanks. I will check out the technical...

RCJK... thanks. I will check out the technical bulletin. We are looking at amplicons in the 550-600 bp range and we are using the Lib-L kits.
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