I am looking to do a project using the yeast KO collection. In the collection are over 6000 deletion strains, with a unique 20mer barcode in each KO. I am planning to use HIseq, and I know I can build the adapters and Illumina barcodes into the PCR primer, because the amplicon length will be the same for each (~120bp). The concern is that of the 120 bases, only the 20mer will be a unique sequence, and a colleague said I should make a 10bp linker in front of the area to be sequenced to help increase library complexity. Someone else said that phiX spikeins at 30% would be sufficient. I am just confused as to what I should do at this point. Hopefully someone has done this and can help me out. I appreciate any help I can get.
-
-
1) have NNNN sequences to introduce diversity for the first 6 bp or so that is required
2) design staggered primers so that a part of your read will start on a different base (have offset versions that start extension @ 1, 2, 3, 4, 5 bp for example) -
Hi dear
I am doing the same project and we are about to use the 20mers as well.
I also have another question. When I add the inhibitory compound to my pool library in liquid pool, I can start with the OD of 0.06 - 0.08. When I go higher , my compound can not inhibit the growth well. I was wondering if you also start this OD of cells of a certain cell density. I have got the diploid library.
Which paper you are following?Comment
-
NGS has revolutionized clinical microbiology by offering new ways to identify and surveil pathogens without prior knowledge of the target. One popular approach is 16S rRNA and ITS amplicon sequencing, which targets specific microbial markers. But studies are increasingly incorporating shotgun metagenomics, a technique that sequences all DNA from a sample without targeting specific microbial markers. This approach allows more detailed taxonomic classification, detection...10-30-2026, 06:46 AM -
Tumors contain complex mixtures of cell populations that evolve in response to treatment and host-specific factors. Tissue biopsies provide only static snapshots of a tumor’s state. But what if we could follow that evolution over time using a minimally invasive technique?
Liquid biopsies can detect tumor-derived material circulating in the bloodstream, providing a way to study treatment response, detect emerging resistance, and discover new biomarkers. Circulating...10-14-2026, 09:55 AM
Comment