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  1. ....5% trypsin and resuspended in fresh media for counting.CUT&TagCUT&Tag was performed according to the EpiCypher CUTANA Direct-to-PCR CUT&Tag protocol using 100,000 cells (details in Supplemental Methods), with a light crosslinking step adapted from a published protocol (Kaya-Okur et al. 2020). All CUT...
  2. .... (C) Number of data sets in which a high-confidence event is significant for expression (left) and splicing (right). (D) Summary of all high-confidence events in the moderate and long alternative exon sets. Euler plot of gene-level overlaps between the two sets is shown at right. (E) 32P-labeled RT-PCR...
  3. ...in pancreatic and other cancer cell lines, we conducted nested PCR and Sanger sequencing (Supplemental Fig. S8D–F). Isoform 2 was validated in Capan-1 and SNU-16 cells but absent in hTERT-HPNE normal cells (Fig. 3E; Supplemental Table S6).View larger version: In this window In a new window Figure 2...
  4. ...binding of a subset of RNA-binding proteins correlates with trans interaction of the encoding loci. We observe that these trans-interacting loci are close to nuclear speckles. These findings support the existence of trans-interacting chromatin domains (TIDs) driven by RNA biogenesis. Trans-C provides...
  5. ...Biosciences). After clonal expansion, genomic DNA was extracted from each clone using the SimplePrep reagent for DNA (Takara, #9180) and screened for mutations by quantitative PCR (qPCR). Candidate clones were further verified for mutations by Sanger sequencing.RNA interferenceSmall interfering RNA (si...
  6. ...of Chromosome 8 following AZ3146 treatment.Following treatment with AZ3146, we isolated 52 viable iPS clones. TaqMan qPCR using 8p-specific probes did not reveal any clones with a euploid karyotype. However, we identified two clones that appeared to have acquired a trisomy of Chromosome 8 (Fig. 2B; Supplemental...
  7. ...Functional genomics analysis of developing zebrafish and human endoderm reveals highly conserved cisregulatory modules acting during vertebrate organogenesis Daniela M. Riley,1,7 Randa Elsayed,1,7 Mark D. Walsh,2,7 Simaran Johal,2 Ying Lin,3,4 Harry Walton,1 Till Bretschneider,5 Sascha Ott,1...
  8. ...Pan analysis reveals families of ubiquitin-ligase adaptors as key genomic divergence drivers that lead to hybrid incompatibility Dongying Xie1,2,3, Pohao Ye1,3, Yiming Ma1 and Zhongying Zhao1 1Department of Biology, Hong Kong Baptist University, Hong Kong SAR, China; 2Institute for Research...
  9. .... Subsequently, the complementary DNA (cDNA) was amplified for 12 PCR cycles. We used AMPure XP bead cleanup (1.8×), followed by size selection using 3% agarose, dye-free gel cassettes with internal standards (Sage Science BDQ3010) on the BluePippin platform. Sequencing was performed on a NovaSeq 6000 platform...
  10. ...using the NEB Ultra II directional RNA library prep kit with sample purification beads (NEB E7765), with 10–14 PCR cycles (determined empirically to achieve the minimum number of cycles per library). Libraries were then sequenced on an Illumina HiSeq 2000 in single-end mode, except for the 12 h time...
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