Library Preparation for NGS of immune repertoire?

Lael Werner @Lael_Werner5

07 July 2019 1 5K Report

Anybody has experience with a kit for library preparation from DNA.

For T and B immune repertoire , for illumina sequencing.

BUT

samples are from FFPE!!

I use Lymphotrak and it is great with fresh tissue or blood, but not FFPE

Benjamin THOHA Thomas

following

Badges
Science topic

More Lael Werner's questions See All

Experience with Foxp3 intracellular staining?

Hello. Need help with (eBioscience) FoxP3 kit. I heard the external antibodies (CD4 and CD25) can be added, AFTER the fix/perm, after overnight incubation at 4C, CONCURRENTLY with the Foxp3...

31 December 2016 2,099 5 View

How much IL-10 secreted from 1million (isolated) CD4+ cells?

How much IL-10 can be expected to secreted from 1million (freshly isolated and magnetically seperated, human) CD4+ cells, stimulated for 24 and/or 48 hours 1ug/ml plated CD3+ 2ug/ml soluble...

11 December 2016 3,983 2 View

Anti-CD3 for stimulation is displaced?

I have been activating T cells with anti-CD3 (either soluble or plate bound) for many years. Currently, I am establishing a protocol whereby at the time of coating (or subsequently) I plate bind...

09 October 2016 2,233 1 View

Can I base on reverse DNA sequences to perform alignment, convert to amino acids and GenBank submission?

I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?

11 August 2024 5,138 1 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

Microtome tissue sectioning - How to prevent chatter of paraffin tissue specimen sections?

Hello everyone, I have recently started using the microtome device for sectioning of paraffin-embedded mice lung. While I had some success in sectioning and observing proper ribbons, some of my...

06 August 2024 666 4 View

Do you have good tips for seaweed tissue preservation in the field for post RNA extraction?

I will be with my students collecting seaweed samples in a marine farm and later we will process this tissue for RNA isolation and further sequencing. Does anyone have tips on how to collect the...

04 August 2024 501 2 View

How to Transform my handy-wavy explanation of why group I/II metals oxides in aqueous solution are basic into a rigorous argument?

I heard an explanation about something being a better proton acceptor or lone pair donor but that doesn't make sense. I couldn't explain in in terms of acid-base theory. The hand-waving way I saw...

03 August 2024 9,918 0 View

What is the problem with these tissue culture plants?

All plants are green but some of these plants becomes yellow. I did not found any reason. Please help me to find out the real problem.

01 August 2024 589 4 View

Does anyone have issues using Prepman Ultra reagent for MicroSeq ID bacterial, fungal and yeast sample preparation?

I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the...

01 August 2024 2,079 0 View

For the process of preparing decellularized matrices what is the ideal solution in which we can bring the tissue from slaughter house to lab?

I am planning to prepare a decellularized porcine tissue derived biomaterial. It would be very helpful if I get to know the composition of the solution which is ideal to bring the tissue sample...

31 July 2024 5,645 2 View

Culture fibroblasts with or without FGF2?

I'm new to primary human fibroblast culture and I'm studying aging using cells from old and young donors. Some research groups have used media supplemented with FGF2, so I cultured cells in the...

29 July 2024 3,030 2 View

While using the IHC PROFILER plugin in image j software, to quantify the epithelial cytoplasmic DAB staining do you crop remove the stromal tissue?

My question pertaining to the DAB staining in cytoplasm of human oral squamous cell carcinoma tissue. When quantifying the epithelial cancer cells do we have to crop remove the stromal tissue?...

29 July 2024 2,682 6 View