2g). Similarly, full-length coverage with Smart-seq3xpress resulted in significantly (2–5-fold) increased read support over exon–exon and exon–intron splice junctions, which form the basis for RNA velocity inference16 (Fig. 2h). To perform a direct comparison of cluster granularity, we ...
cDNA synthesis mixture was prepared by combining 5.5 μl DEPC-treated water, 4 μl of 5x reverse transcriptase buffer (Fermentas, St. Leon-Rot, Germany), 2 μl of dNTP (10 mM), 5 μl of DNase treated RNA (about 2 μg) and 2.5 μl of 40 μM Cy5-labeled anchored poly(dT)-...
The first step in this process consists of the cDNA synthesis in which a DNA complementary to the RNA is synthesized with the help of a reverse transcriptase enzyme in the presence of dNTP and DNA primers. An oligo dT primer is used to make cDNA of poly A-RNA such as mRNA. In ...
Microbial communities are responsible for biological wastewater treatment, but our knowledge of their diversity and function is still poor. Here, we sequence more than 5 million high-quality, full-length 16S rRNA gene sequences from 740 wastewater treatment plants (WWTPs) across the world and use ...
Here we describe an improved form of the AID system. Depletion of Mcm4-aid protein was greatly enhanced by expression of Skp1-TIR1, a fusion protein comprising plant TIR1 and fission yeast Skp1. This i-AID system (i mproved AID system) was applicable for depletion of other essential ...
Artemis first, before ligation. Consistent with the critical role of DNA-PKcs in end-processing, DNA-PKcs null lymphocytes accumulate hairpin CEs, leading to T- and B- SCID in mouse models [49,50,51]. Meanwhile, DNA-PKcs null cells can ligate SEs to form signal joints (SJs) effectively ...
For example, in breast cancer tissue: (1) the “sulfatase pathway” and “aromatase pathway” are upregulated, thereby providing a steady stream of precursors for estrogen synthesis in the form of inactive steroid sulfates or androgens, respectively, and (2) the reductive 17-β-hydroxysteroid ...
1. Prepare the reaction mixture to have the total volume of 50 l by combining the following reagents. Template 10x LA PCR Buffer II (Mg2+ plus) (supplied with LA PCR Kit Ver.2) dNTP Mixture (2.5 mM) S1 Primer (20 pmol/l) A1 Primer (20 pmol/l) TaKaRa LA Taq (5 units/l) ...
The reaction mixture contained 0.1 mM dNTP, 0.5 mM of each degenerate primer and 1.0 U of HiFi-Taq DNA polymerase (TransGen Biotech, Guangzhou, China) in a total volume of 25 μl. The first PCR was carried out with the following conditions: initial preheating for 5 min at 94°C, 35 ...
All PCR reactions were carried out in a 10 μL volume with a final concentration of 1x PCR buffer, 1.5 mM MgCl2, 200 μM of each dNTP and 0.1 μM of each primer, 10 ng genomic DNA, and 0.5U of Platinum®Taq DNA Polymerase (Invitrogen TermoFisher). The amplification profile was: ...