Journal: Nature Protocols

Article Title: Highly multiplexed targeted DNA sequencing from single nuclei

doi: 10.1038/nprot.2016.005

Figure Lengend Snippet: Figure 1 | Protocol overview. Step 1: tissue or cells are lysed and nuclear suspensions are prepared and stained with DAPI. Steps 2–9: nuclei are flow-sorted using FACS by gating the G1/0 or G2/M distributions based on total DNA content, and single nuclei are deposited into individual wells in a 96-well plate. Steps 10 and 11: the nuclear membrane is lysed. Steps 12 and 13: WGA using MDA is performed using the Φ29 polymerase. Steps 14–16: for quality control of WGA efficiency, each WGA reaction is screened with a 22-amplicon qPCR panel using chromosome-specific primer pairs. Steps 17–61: barcoded libraries are prepared from each single-cell WGA reaction. Steps 62–103: barcoded libraries (12–96) are pooled together into a single reaction followed by targeted capture. Step 104: the pooled libraries are then used for NGS on the Illumina platform.

Article Snippet: 29 Polymerase (10 units per 1 μl; NEB, cat. no. M0269l; Polymerase buffer is included) dNTP set, 100 mM (NEB, cat. no. N0446S) PBS 1× Qubit dsDNA high-sensitivity assay kit (Invitrogen, cat. no. Q32854) ! cautIon The dye is light sensitive.

Techniques: Staining, Membrane, Control, Amplification

Journal: eLife

Article Title: Channel nuclear pore complex subunits are required for transposon silencing in Drosophila

doi: 10.7554/eLife.66321

Figure Lengend Snippet:

Article Snippet: Commercial assay or kit , Deoxynucleotide Solution Set (100 mM; 25 μmol each) , New England Biolabs , Cat# N0446S , .

Techniques: Magnetic Beads, Lysis, Protease Inhibitor, Transfection, Amplification, Recombinant, Purification, Software