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.
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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
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The immune system’s power comes from its genetic diversity, allowing myriad threats to be neutralized through first recognizing foreign antigens. That diversity is also what makes the immune system so difficult to study. Recent advances in sequencing technology and computational biology, however, are giving researchers new tools to understand immune responses and immune-related diseases in greater detail.
This convergence of genetics, immunology, and computation...09-01-2026, 05:41 AM
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