ASHG: Primary Template-directed Amplification 2021年3月9日发布 03:53 ASHG: Primary Template-directed Amplification 讨论 登录参与讨论 这里的评论内容走失了 请检查网络后,点击空白处重试特色推荐 杀毒软件 软件下载 手机版 Windows版 Mac版 iPad版 TV版 服务 客服 反馈 侵权投诉 VIP采购 腾讯视频隐私保护...
Primary template-directed amplification (PTA) is an improved amplification technique for single-cell DNA sequencing. We generated whole-genome analysis of 76 single neurons and developed SCAN2, a computational method to accurately identify both clonal and non-clonal somatic (i.e., limited to a ...
PCR amplification was performed with Phusion or Q5 High Fidelity DNA Polymerase (New England Biolabs). Primer sequences are provided in Supplementary Table 4. On and off-target genome editing analysis PCR products were purified using the NucleoSpin Gel and PCR Clean-up kit (Machery-Nagel). ...
Enhancing CRISPR-mediated site-specific transgene insertion efficiency by homology-directed repair (HDR) using high concentrations of double-stranded DNA (dsDNA) with Cas9 target sequences (CTSs) can be toxic to primary cells. Here, we develop single-str
Variable heavy and light chains were synthesized using a modified SMARTSeq-V4 protocol by 5' RACE. Single-cell RNA was first purified with RNAclean beads (Beckman Coulter). cDNA was then synthesized using 5' RACE (rapid amplification of cDNA ends), adding distinct 3' and 5' template switch ...
Genomic alterations were identified at osimertinib progression (liquid biopsy in blood or pleural fluid) considering mutations (A) and amplifications (B). All patients in cohort A had two or more commutations, while in cohort B, only three had a commutation.CAmplifications identified at the time...
Primers for Ki-67 amplification were designed in our laboratory as described previously [23]. Expression of β-ACTIN (forward: 5′-TCTACAATGAGCTGCG-3′ and reverse: 5′-AGGTAGTCAGCTAGGT-3′) was used as a reference housekeeping gene. All primers were run through the National Center for...
In the negative control, ddH2O was used instead of the cDNA template. Western Blot Brain samples were lysed with RIPA lysis buffer (50 mM Tris pH 7.5, 150 mM NaCl, 1% Triton X-100, 1% Nadeoxycholate, 0.1% SDS and Halt proteinase inhibitor [Pierce, USA; No. 78430]). Total protein ...
Engineering T cell specificity and function at multiple loci can generate more effective cellular therapies, but current manufacturing methods produce heterogenous mixtures of partially engineered cells. Here we develop a one-step process to enrich unlabeled cells containing knock-ins at multiple target lo...
The first cDNA amplification (18–20 cycles) was followed by cDNA fragmentation and isolation of 300–500 bp long fragments. After ligation of the 39 Illumina adapter, a final PCR (4 cycles) was performed. For the preparation of the non-enriched transcriptome (RNA-seq, all transcripts ...