Forum: Sample Prep / Library Generation
05-27-2020, 06:44 AM
|
|
Replies: 0
Views: 1,429
RNA-Seq on "real" single fixed cells?
Hi,
Anyone with experience or advise on how to perform RNA-Seq (mRNA, total RNA,...) on "real" single fixed cells? So not thousands of individual cells but on individual cells from a collection of...
|
|
Forum: Sample Prep / Library Generation
02-06-2020, 04:24 AM
|
|
Replies: 3
Views: 1,381
|
|
Forum: Sample Prep / Library Generation
01-17-2020, 04:21 AM
|
|
Replies: 0
Views: 1,531
Nextera DNA Flex and WGA
Hi,
Anyone tried to apply Nextera DNA Flex (Illumina) to WGA of human single cells?
Are using Picoplex Gold (Takara, WGA followed by PCR to add tails to make the WGA into a NGS library) but would...
|
|
Forum: Illumina/Solexa
04-16-2019, 01:09 AM
|
|
Replies: 19
Views: 8,919
I am working in a hospital lab and am the happy...
I am working in a hospital lab and am the happy user of two Sciclone NGS, a NGS Express and a Zephyr NGS. We use the robots for DNA cleanup, NGS lib prep, purification and QC.
There are a number of...
|
|
Forum: Illumina/Solexa
03-29-2019, 03:01 AM
|
|
Replies: 2
Views: 733
I have received the following clarifying reply...
I have received the following clarifying reply from Illumina Techsupport:
"The run number assignment is sequential to which side actually starts first after the pre-run checks. Usually FCA will have...
|
|
Forum: Illumina/Solexa
03-25-2019, 03:05 AM
|
|
Replies: 2
Views: 733
|
|
Forum: Sample Prep / Library Generation
03-11-2019, 04:59 AM
|
|
Replies: 2
Views: 1,426
Hi Arctan,
We have also done some bacterial...
Hi Arctan,
We have also done some bacterial RNA-Seq using Ribo-Zero (and ScriptSeq) and have observed the same peak. I have "only" generated the data, but have not received any feedback from the...
|
|
Forum: Sample Prep / Library Generation
01-30-2019, 07:01 AM
|
|
Replies: 17
Views: 8,263
"To check if the large fragments are actually...
"To check if the large fragments are actually overamplification?"
Two tests to do:
1) denature a small aliquote of the library and run it on e.g. a Bioanalyzer. If the large peak is due to...
|
|
Forum: Illumina/Solexa
01-18-2019, 02:18 AM
|
|
Replies: 9
Views: 3,337
In our hands, we do not find any significant...
In our hands, we do not find any significant differences in the quality between insert data with perfect index reads and insert data with 1-mismatch index reads.
So despite a much larger fraction of...
|
|
Forum: Illumina/Solexa
01-15-2019, 05:12 AM
|
|
Replies: 9
Views: 3,337
|
|
Forum: Illumina/Solexa
01-15-2019, 05:03 AM
|
|
Replies: 9
Views: 3,337
|
|
Forum: Sample Prep / Library Generation
01-10-2019, 02:01 AM
|
|
Replies: 2
Views: 2,139
Take a look at...
Take a look at https://ls.beckmancoulter.co.jp/files/appli_note/Gel_Free_Using_SPRIselect.pdf
But I agree with nucacidhunter that 50 bp is a too narrow window for a double-SPRI
|
|
Forum: Illumina/Solexa
12-20-2018, 01:24 AM
|
|
Replies: 9
Views: 3,337
|
|
Forum: Illumina/Solexa
12-19-2018, 02:57 AM
|
|
Replies: 9
Views: 3,337
|
|
Forum: Sample Prep / Library Generation
08-27-2018, 02:08 AM
|
|
Replies: 4
Views: 2,034
|
|
Forum: Illumina/Solexa
07-25-2018, 12:21 AM
|
|
Replies: 6
Views: 2,238
We have been using Ribo-Zero + ScriptSeq for...
We have been using Ribo-Zero + ScriptSeq for years with success. Primarily for samples of human RNA (using Ribo-Zero GOLD or Globin-GOLD) but also a few samples of bacterial RNA (of course using the...
|
|
Forum: Sample Prep / Library Generation
05-18-2018, 03:03 AM
|
|
Replies: 0
Views: 1,599
gDNA library prep kit for NovaSeq 6000?
Going to have a NovaSeq installed soon and wonder which library prep kits are most optimal for human gDNA? We are going to use the libraries for both WGS and for capture (exome/panels). Index hoping...
|
|
Forum: Illumina/Solexa
11-22-2017, 11:08 PM
|
|
Replies: 12
Views: 3,177
A update on the chemistry-only recipe....
We...
A update on the chemistry-only recipe....
We got a custom recipe from Illumina (without any guarantee) but it didn't work. It was supposed to do chemistry only in cycle 13-17 - but it didn’t.
/Jakob
|
|
Forum: Illumina/Solexa
10-26-2017, 02:14 AM
|
|
Replies: 13
Views: 2,637
|
|
Forum: Sample Prep / Library Generation
10-23-2017, 06:42 AM
|
|
Replies: 1
Views: 1,217
Hi,
If we happen to have libraries of low...
Hi,
If we happen to have libraries of low concentration, we denature with 0.2 M NaOH, add an equal volume of 200 mM Tris-HCl, pH 7, and adds HT1 to reach 20 pM.
An example: a library is 230 pM....
|
|
Forum: Illumina/Solexa
10-20-2017, 12:34 AM
|
|
Replies: 2
Views: 1,346
Hi kerryp,
No problem. Have done it...
Hi kerryp,
No problem. Have done it successfully for several years doing Nextera XT libraries of bacteria and sequencing on NextSeq medium in batches of 50-70 bacteria.
I havent had concerns about...
|
|
Forum: Illumina/Solexa
10-19-2017, 12:57 AM
|
|
Replies: 12
Views: 3,177
|
|
Forum: Illumina/Solexa
10-18-2017, 11:14 PM
|
|
Replies: 12
Views: 3,177
Hi Provirus and Austinso,
What a coincidence....
Hi Provirus and Austinso,
What a coincidence. We have just been in contact with Illuminas Techsupport to have a custom program made for the NextSeq 500 to do a PE run with 5 dark cycles after the...
|
|
Forum: Illumina/Solexa
08-22-2016, 11:45 PM
|
|
Replies: 2
Views: 4,606
Hi, thank you very much for sharing this.
I...
Hi, thank you very much for sharing this.
I have made a simple and a bit rough R-script (attached) to compare two list of indexes and return a similarity matrix, which can be used to select index...
|
|
Forum: Sample Prep / Library Generation
07-27-2016, 02:36 AM
|
|
Replies: 0
Views: 976
LabChip GX - daisy chains?
Hi,
We are happy using a LabChip GX for our NGS work - both QC of gDNA and of libraries.
But we have an issue with daisy-chains in NGS libraries which seem to be related to the GX and not the...
|
