Volume 12 Issue 4, April 2017

Ståhlberg et al. describe SiMSen-seq, a barcoding NGS library preparation approach to enable detection of rare variant alleles from as little as 5 ng of input DNA. The method also works for fragmented templates such as cell-free DNA.

This protocol describes how to grow a functional and transplantable corneal epithelium and how to generate ocular-like cell lineages resembling neuroectoderm, neural crest, ocular-surface ectoderm, or surface ectoderm derived from human iPS cells.

Translation complex profile sequencing (TCP-seq) is a major technological step forward for analysis of mRNA translation. Unlike ribosome profiling, TCP-seq can be used to study all stages of translation, from initiation through to ribosome recycling.

This protocol describes how to create a decerebrate mouse model for investigating sensory-motor integration and motor output from the spinal cord.

This protocol describes how to express and purify a protein probe and subsequently conjugate the probe to dyes with different fluorescence properties. The probes have applications in super-resolution imaging.

Solid-state NMR is useful for getting structural information for insoluble proteins. In this protocol, chemical shifts for backbone atoms are assigned using proton detection and ultrafast magic-angle spinning of perdeuterated proteins.

Many cellular processes rely on cells generating or responding to nanoscale mechanical forces. This protocol describes STED–traction force microscopy (STFM), which allows these forces to be measured with higher resolution and accuracy than standard TFM.

Ion-mobility spectrometry (IMS) separates molecules according to charge, shape and size. In this protocol, traveling-wave ion mobility mass spectrometry (TWIMS) is coupled with LC for metabolomics and lipidomics analysis.

This protocol describes how to recapitulate biliary development by differentiation of hPSCs into endoderm, foregut progenitor cells, hepatoblasts, cholangiocyte progenitors and mature 3D cholangiocyte-like cell organoids.

This protocol from Feng Zhang's lab enables genome-scale knockout and transcriptional activation screening using the CRISPR-Cas9 system, as sgRNA libraries are constructed and packaged into lentiviral vectors for delivery into cells for screening.