Three protocol PDFs attached that I stumbled upon on the web, I thought some of you may be interested.
Header Leaderboard Ad
Collapse
454 Library / Emulsion / Instrument Protocols (October 2008)
Collapse
Announcement
Collapse
No announcement yet.
This is a sticky topic.
X
X
-
cDNA from Evrogen with Ti
Has anybody had any experience with Ti sequencing Evrogen Trimmer Direct cDNA? For a 2 region Ti, we got 67 Mb and only 20% pass. The support people feel it is a problem in the emulsion PCR. Any help would be much appreciated. We've sequenced ~30-40 similar samples with FLX and no problems (over 100 Mb, ~440,000 reads per run).
Comment
-
Hello, I´m a new member.
Originally posted by ECO View PostThree protocol PDFs attached that I stumbled upon on the web, I thought some of you may be interested.
Thank you for the three protocol pdf!Last edited by futuro_cortina; 03-09-2009, 11:21 AM.
Comment
-
PTS Wisdom?
Hi
We are currently working on de novo RNA-seq and re-sequencing in a pre-genome species (onion) and are interested in PTS or a similar generic barcoding approach to enable us to do population genetic studies.
If anyone can give me any opinion or advice concerning this approach I'd be really grateful. To date we are just working with barcoded primers
Regards
John
Comment
-
Hi
We are trying to work out cDNA sequencing using 454, but Titanium did not work, and Standard was severely affected by polyA tail despite the nice profile of the sstDNA library.
We used Dynabeads for mRNA purification, and Stratagen Just cDNA kit for ds cDNA synthesis (1st strand primed by none-anchored oligo dT from the kit). Our samples tend to have long polyA tail, and we found 454 sequencing cannot tolerate this.
We are trying anchored dT primer now, but would appreciate any suggestion, such as choice of kit, 1st strand priming method (dT vs random primer?), or any tips in improving library constrcution for greater chance of success in sequencing?
Thank you.
Comment
-
-
re Poly A etc
We have had very nice results from Ti sequencing of normalised cDNA prepared using Evrogen Mint + Trimmer kits, using a modified 3' Primer XXXXXXXXXXX(T)4G(T)9C(T)10VN as described by Beldade et al.
The information posted at http://www.cees.uio.no/research/faci...equencing.html was very useful.
Comment
Latest Articles
Collapse
-
by seqadmin
Developments in sequencing technologies and methodologies have transformed the field of epigenetics, giving researchers a better way to understand the complex world of gene regulation and heritable modifications. This article explores some of the diverse sequencing methods employed in the study of epigenetics, ranging from classic techniques to cutting-edge innovations while providing a brief overview of their processes, applications, and advances.
Methylation Detect...-
Channel: Articles
Yesterday, 10:46 AM -
-
Differential Expression and Data Visualization: Recommended Tools for Next-Level Sequencing Analysisby seqadmin
After covering QC and alignment tools in the first segment and variant analysis and genome assembly in the second segment, we’re wrapping up with a discussion about tools for differential gene expression analysis and data visualization. In this article, we include recommendations from the following experts: Dr. Mark Ziemann, Senior Lecturer in Biotechnology and Bioinformatics, Deakin University; Dr. Medhat Mahmoud Postdoctoral Research Fellow at Baylor College of Medicine;...-
Channel: Articles
05-23-2023, 12:26 PM -
-
by seqadmin
Continuing from our previous article, we share variant analysis and genome assembly tools recommended by our experts Dr. Medhat Mahmoud, Postdoctoral Research Fellow at Baylor College of Medicine, and Dr. Ming "Tommy" Tang, Director of Computational Biology at Immunitas and author of From Cell Line to Command Line.
Variant detection and analysis tools
Mahmoud classifies variant detection work into two main groups: short variants (<50...-
Channel: Articles
05-19-2023, 10:03 AM -
ad_right_rmr
Collapse
News
Collapse
Topics | Statistics | Last Post | ||
---|---|---|---|---|
Unveiling Genetic Associations Through Transcription Factor Binding Quantitative Trait Loci
by seqadmin
Started by seqadmin, Yesterday, 07:50 AM
|
0 responses
4 views
0 likes
|
Last Post
by seqadmin
Yesterday, 07:50 AM
|
||
Exploring French-Canadian Ancestry: Insights into Migration, Settlement Patterns, and Genetic Structure
by seqadmin
Started by seqadmin, 05-26-2023, 09:22 AM
|
0 responses
8 views
0 likes
|
Last Post
by seqadmin
05-26-2023, 09:22 AM
|
||
Started by seqadmin, 05-24-2023, 09:49 AM
|
0 responses
20 views
0 likes
|
Last Post
by seqadmin
05-24-2023, 09:49 AM
|
||
Introducing ProtVar: A Web Tool for Contextualizing and Interpreting Human Missense Variation in Proteins
by seqadmin
Started by seqadmin, 05-23-2023, 07:14 AM
|
0 responses
31 views
0 likes
|
Last Post
by seqadmin
05-23-2023, 07:14 AM
|
Comment