Hello,
I want to sequence enriched RNA (it will include both mRNAs and bacterial small non-coding RNAs (<200 nt)). I would like to fragment...
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Effects of total RNA fragmentation on small RNAs
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quality check 1 ng DNA
Hi everybody
I am willing to sequence with MiSeq using a Nextera XT library starting with very challenging samples of not cultivable bacteria.... -
Weird distribution of insert size
I got a strange distribution of insert size as analyzed by Qualimap on the BAM file. Not totally aware of the library preparation procedure yet, but this... -
RNA-seq of long RNAs (paired-end, stranded) -- fragmentation time
I'm using the TruSeq Stranded Total RNA library preparation kit to generate libraries for 2x150 bp paired-end sequencing. Illumina's fragmentation protocol... -
Stingray mRNA Fragmentation NEBNext
Hi all,
I am using the NEBNext Ultra Directional RNA Library Prep Kit for Illumina following mRNA isolation using dT beads.
... -
Inconsistent size distribution of fragmented mRNA and cDNA for RNA 3Seq
Hello everyone,
I am new to the RNA 3' Sequencing experiment using a protocol developed in house based on Illumina TruSeq mRNA sample prep.... -
fragmentation of ovine DNA
Hi,
this is the first time I will construct a DNA library to next-generation sequencing. I'm working with ovine DNA and trying to set up... -
Incomplete shearing with Bioruptor?
Hi all!
Shearing gDNA for NGS sample prep. I'm trying to shear my DNA into 500bp fragments using the bioruptor and have two problems: ... -
RNA integrity after SpeedVac
Any insight on what happens to RNA in NF water when it's speedvac'd at high heat (~70C)? I'm trying to figure out if it can cause fragmentation. -
Differences between sonicators?
Hi
I'm new here and looking for advice on fragmenting chromatin. We plan to do quite a bit of ChIPseq to identify transcription factor... -
cDNA fragmentation
For RNA-seq libraries, rather than chemically fragmenting mRNA has anyone tried fragmenting 2nd strand cDNA?
Just curious. -
mRNA fragmentation for SPRIworks
We are making ds-cDNA for automated mRNA-seq library preparation using the SPRIworks robot. After the poly-A selection, our protocol calls for fragmentation...Last edited by jowenMIT; 01-27-2011, 02:23 PM. -
Sonicator
Hi all,
We are not quite ready to invest in a Bioruptor and wanted to know what sonicators people are using out there. We've been using a collaborators... -
New fragmentation methods
Physical shearing methods for double stranded DNA include sonication, nebulization and hydroshearing. We use all three. They work well for constructing... -
RNA-Seq: cDNA prep and fragmentation questions
I'm developing libraries for RNA-seq on the Illumina platform. I have a couple of questions:
1.) After you have made double stranded cDNA...
