Adapterama Info & Protocols

Dissolving iTru Primers

Handling iTru primers in plates (1.25 nmol / well; from us).

Making iTru Y-yoke adapter stubs

Making iTru Y-yoke adapter stubs (5nmol aliquots; from us).

Making iTru Kapa libraries.

Making iTru libraries with Kapa reagents.

Making iNext libraries.

Making iNext libraries with NEB & Kapa reagents.

iTru 3RAD Libraries No Molecular IDs

Dual-digest RADseq libraries with quadruple indexing.

Making Adapters from Oligos

How to create double-stranded adapters from oligos (primers) that you order yourself (not from us).

Speedbead Recipe and Directions

How to make, use, and validate Speedbeads.

3RAD with Molecular IDs

Dual-digest RADseq libraries with PCR duplicate tagging.

3RAD Adapter Designs

Dual-digest RADseq adapters - 4 x 2 designs.

Quickest 3RADs

Dual-digest RADseq libraries with quadruple indexing.

Making iTru Amplicon libraries.

Making iTru libraries for PCR products with NEB reagents.

Dissolving iTru Blocking Oligos

How to dissolve iTru blocking primers to make adapter blockers.

Dissolving and Annealing 3RAD Adapters

How to create double-stranded adapters from normalized oligo pairs (what we send out).

UCE Protocols

On-bead Illumina Libraries

This was a big deal at the time, but you can now do even better with Kapa Hyper Prep Kits.

UCE Sequence Capture with Universal Blockers

Again, this was helpful at the time, but you can now do better by using the latest MYbaits-manual from Arbor Biosciences (formerly Mycroarray).

Post-enrichment limited cycle PCR

This can be good to have, but you can simply use the latest MYbaits-manual from Arbor Biosciences (formerly Mycroarray).

Validations of enrichments via qPCR

This is a reasonable way to test if new baits are doing what you want before you spend a lot on sequencing. However, now that we have so many iTru primers, it will usually be cheaper and better to accomplish this task via low depth sequencing.