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Forum: Illumina/Solexa 05-07-2017, 11:09 AM
Replies: 1
Views: 1,053
Posted By soleulloa
46% reads >Q30 in R2... Why? 😔

Hi all,

I've used a MiSeq 500 cycles v2 and 10 pM of my libraries pool + phiX 10%.
Everything was ok but at cycle 491 the run was stopped (I don't know why).
I was looking for some paramethers...
Forum: Illumina/Solexa 02-17-2017, 09:27 AM
Replies: 11
Views: 1,839
Posted By soleulloa
Someone has quantified libraries before dilution?...

Someone has quantified libraries before dilution? When you prepare 2 nM dilutions?
I've quantified before and after that final dilution now because this is the first run when I have this problem and...
Forum: Illumina/Solexa 02-16-2017, 06:15 AM
Replies: 11
Views: 1,839
Posted By soleulloa
Hi, 95.27% PF

Hi,

95.27% PF
Forum: Illumina/Solexa 02-16-2017, 04:02 AM
Replies: 11
Views: 1,839
Posted By soleulloa
Ok! Thank you. I'm using 10 pM (from 4 nM) to...

Ok! Thank you.
I'm using 10 pM (from 4 nM) to get this result.
What do you think I have to use now? 12 or 15 pM to get a good run?
Forum: Illumina/Solexa 02-15-2017, 01:14 PM
Replies: 11
Views: 1,839
Posted By soleulloa
Discordance between Cluster Density and Thumbnail images

Hi all,

I ran MiSeq using a 500 cycles kit.
The value for cluster density was 310K/mm2 but if I'm going to look the thumbnail images, I think I have much more cluster than that.

Illumina tech...
Forum: Illumina/Solexa 12-08-2016, 06:32 PM
Replies: 6
Views: 1,654
Posted By soleulloa
Thanks all. My question is relative about "bad...

Thanks all.
My question is relative about "bad quality assembly".
How many contigs less than 1000 bp do you expect in a MiSeq 300 cycles run? How much of those are ok?
I am trying to link that...
Forum: Illumina/Solexa 12-07-2016, 09:13 AM
Replies: 6
Views: 1,654
Posted By soleulloa
Is there any relation between DNA integrity and contigs size?

Hi all,

I need to know if DNA integrity has any relation with contigs size?
For example, if my DNA is more degraded I should obtain littlest contigs?

Thanks
Soledad
Forum: Illumina/Solexa 06-13-2016, 12:43 PM
Replies: 10
Views: 1,834
Posted By soleulloa
Hi all, I've contacted Illumina support...

Hi all,

I've contacted Illumina support immediatly. They ask for me some archives (sample sheet, runinfo, runparameter and InterOp) and finally the answer was:
YOUR SAMPLES ARE VERY CONCENTRATED...
Forum: Illumina/Solexa 06-13-2016, 05:48 AM
Replies: 10
Views: 1,834
Posted By soleulloa
Pleeeease.. any of you have seen that error...

Pleeeease.. any of you have seen that error before?
I'm afraid to run again and lose the reagents!
:(
Forum: Illumina/Solexa 06-10-2016, 09:58 PM
Replies: 10
Views: 1,834
Posted By soleulloa
Hi kmcarr, Why do you think this is a hardware...

Hi kmcarr,
Why do you think this is a hardware error? Have you seen it before? ��
Forum: Illumina/Solexa 06-10-2016, 12:17 PM
Replies: 10
Views: 1,834
Posted By soleulloa
Z Motor: ADC noise which exceeds ADCNoiseErrorLimit

Hi all,
Yesterday a run in MiSeq was stopped showing the next message:

Z Motor: ADC noise is 3592 which exceeds ADCNoiseErrorLimit 3277. numSamples=10x8, avg=31258, posNoise=1906, negNoise=3592
...
Forum: Illumina/Solexa 10-22-2015, 04:33 AM
Replies: 0
Views: 1,054
Posted By soleulloa
Tween 20% affects Flow cell ??

Hi,
I have some problems in pre-check of a run and MiSeq can't initiate correctly by a flow rate error.
I put reagents at 4C and I did a wash using H2O+Tween 20% for clearing lines, but I forgot...
Forum: Illumina/Solexa 08-20-2015, 08:46 AM
Replies: 5
Views: 5,515
Posted By soleulloa
Thank you so much Jessica! I'll take your...

Thank you so much Jessica!
I'll take your advices :)
Forum: Illumina/Solexa 08-19-2015, 07:44 AM
Replies: 5
Views: 5,515
Posted By soleulloa
Hi Jessica D. Do you use 2nM libraries always...

Hi Jessica D.
Do you use 2nM libraries always to work? And what is the cluster density that you obtain routinely?
Because I've working using 4nM libraries previous to denaturation and I've got...
Forum: Illumina/Solexa 05-18-2015, 07:32 AM
Replies: 30
Views: 8,506
Posted By soleulloa
Thanks Phillip!!! I need to understand how...

Thanks Phillip!!!

I need to understand how much phiX (in percentage) I have in my final pool (including phiX spike in). For example, in my first run I did exactly I described above for spike-in...
Forum: Illumina/Solexa 05-15-2015, 07:08 PM
Replies: 30
Views: 8,506
Posted By soleulloa
Red face Calculating phiX concentration...

Hi all!
I'm a new user of MiSeq and I'm confused about phiX concentrarion to spike in.
In Nextera XT DNA protocol they show a list of dilutions to prepare a final solution of phiX 12,5 pM. Then you...
Forum: Illumina/Solexa 05-15-2015, 06:52 PM
Replies: 4
Views: 2,215
Posted By soleulloa
Hi docsascha! Did you tried a new run?? Tell us...

Hi docsascha!
Did you tried a new run?? Tell us the results! I'm interested in mycobacterium sequencing in my lab :)
Regards
Forum: Ion Torrent 09-04-2014, 10:05 AM
Replies: 0
Views: 1,303
Posted By soleulloa
Is there relationship between library preparation and low quality data?

Hi all!

If:
- all the paramethers about the initial DNA are OK (quantity, 260/280, 260/230 and integrity)
- Bioanalyzer results are OK
- library post OT2 is OK
- run results are OK (% loading...
Forum: Ion Torrent 01-13-2014, 07:07 AM
Replies: 3
Views: 2,802
Posted By soleulloa
Exclamation Wrong library size reads

Hi all,

I've prepared a library using the protocol for 200 bp reads and checked by Bioanalyzer library size was 238 bp.
Then, OT runs and PGM runs, but the readings obtained were too short (53...
Forum: Ion Torrent 10-29-2012, 06:37 AM
Replies: 3
Views: 2,229
Posted By soleulloa
Exclamation Average fragment length post-sonication

Hi all,

I have a question! :)
Anyone knows what is the average fragment length obtained post-sonication when you are preparing a fragment library (from genomic DNA).

For example, using GS...
Forum: Ion Torrent 03-30-2012, 06:31 AM
Replies: 1
Views: 2,882
Posted By soleulloa
Lightbulb Barcoding.... a lot of questions!!!!

Hi all,

I'm Soledad from Chile :)
I want to do amplicon sequencing, but I don't understand very well the neccesary kits.
My amplicons are 1400 bp length, so I think that I have to shear to make...
Forum: Ion Torrent 11-17-2011, 07:31 AM
Replies: 15
Views: 6,654
Posted By soleulloa
Wowwwwww thank you so much!!!! :D

Wowwwwww thank you so much!!!!

:D
Forum: Ion Torrent 11-17-2011, 07:02 AM
Replies: 15
Views: 6,654
Posted By soleulloa
Hi nickloman, Thanks for your early...

Hi nickloman,

Thanks for your early response!
I've being looking for Ion Community, but I can't find information about this :(

I need someone that was performed 316 runs... pleaaaaase!!!
Forum: Ion Torrent 11-17-2011, 06:47 AM
Replies: 15
Views: 6,654
Posted By soleulloa
Hi all, I'm preparing our first run using...

Hi all,

I'm preparing our first run using 316, but I'm not sure if I have to use the same "user guide" that we used for 314 preparation.
Is there any changes in protocols for 316?
Please, help...
Forum: SOLiD 09-20-2011, 12:02 PM
Replies: 3
Views: 2,014
Posted By soleulloa
Thank you Phillip!!! I'll try to do using more...

Thank you Phillip!!! I'll try to do using more time for nick translation ;)
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