4.3. Sequencing Cell suspensions obtained from eight fractions were used for plasmid isolation followed by PCR amplification with the primer pair 5′-GTGCTCAAGCTTTAATACGACTCACTATAGGG-3′ and 5′-GCTCTCGTCCGTCTCTTTC-3′. The PCR products were purified and checked with electrophoresis in 2% agaro...
forward 5′-AGCTACCATG CCCTATGTGG-3′—reverse 5′-ATCAGCACTC CAAACCCAAC-3′; on the other strand, h_SLC6A3_Rev PyroMark Custom Assay—forward 5′-AGGTGGAGGT TTTAATAGGTAAT-3′—reverse [Biotin] 5′-AACCACATTT TACTATATAAACCCA-3′—All the methodological pyro-sequencing details are ...
Sequencing of a single-cell genome requires DNA amplification, a process prone to introducing bias and errors into the amplified genome. Here we introduce a novel multiple displacement amplification (MDA) method based on the unique DNA primase features ofThermus thermophilus(Tth) PrimPol.TthPrimPol di...
Biotinylated PCR products were then processed using the PyroMark Q24 Vacuum Workstation (Qiagen, Hilden, Germany), and subsequent pyrosequencing was performed on a PyroMark Q24 pyrosequencer with PyroMark Gold Q24 Reagents (Qiagen, Hilden, Germany) and the following sequencing primers: CSP-3F (...
Dideoxy sequencing reactions were conducted on the same DNA template used for the in vitro transcription of the UTR240 constructs, using the Thermo Sequenase Cycle Sequencing Kit (Affymetrix) and the same NEDTM-labeled primers as those used for primer extension (Table S1). Sequencing reactions con...
(CSP) sequence fromC. cajan. CSP was amplified using specific primers from theC. cajancDNA sample, cloned into pGEMT-easy vector and confirmed by Sanger sequencing. Similar toCpF3′5′H_2, the confirmed CSP sequence was sub-cloned into the 5′ region of mGFP in pCambia1302 vector by ...
The basic assay includes the following steps: (1) Minigenes construction; (2) in silico analysis; (3) site-directed mutagenesis; (4) transfection of the wild type and mutant minigenes; (5) inhibition of nonsense-mediated decay and RNA purification; (6) transcript sequencing and fragment ...
pre-designed TaqMan gene expression primer/probes assay against the genes GATA4, MEF2C, TBX5, HAND2, NKX2.5, and GAPDH. Real-time quantification system Applied Biosystems 7900 Fast Real-Time PCR system was used for the experiment. The TaqMan gene expression assay IDs of the respective target ...