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To identify DNA-preparations with a low percentage of recombined plasmid we performed PCRs on serial dilutions of template DNA (100 pg–0.01 fg) using primer pairs that detected recombination (key resource table). DNA preparations with the smallest fraction of recombined plasmids were selected for...
Following removal of rRNA using the Ribo-Zero™ Magnetic Gold Kit, the isolated mRNA was fragmented (200–500 nt) by adding fragmentation buffer. First-strand cDNA was then generated by reverse transcription using the isolated mRNA as a template and random hexamer primers. Second-strand cDNA w...
No template control samples, dissociation curves and samples containing the untranscribed RNA were measured in parallel as controls. Data were analyzed with the comparative CT method and presented as 2−ΔCT or 2−ΔΔCT as described77. Constitutively expressed HPRT was measured for internal ...
R-loops primarily form co-transcriptionally (in cis), when the nascent RNA hybridizes back to the DNA template strand [33], but may also form in trans using the homologous recombination (HR) factors Rad52 and Rad51 [34] (Figure 2(b)). Given that the key HR protein in yeast, Rad52...
A template on how short defini- tions should be written has formalized by a dedicated WHO team in 2014 only [14]. On the whole, there appears in many TAGs to have been an understandable focus on medical issues but at the expense of those related to organizing knowledge into an information...
[20]. TERC serves as a template for the synthesis of telomeric repeats and acts as a scaffold that brings the protein subunits of telomerase together with other accessory proteins associated with the complex. Additionally, TERC was shown to have a catalytic function in the process of adding ...
(Clontech, USA) was used to synthesize cDNA with mRNA as template. PCR was used to amplify the cDNAs. cDNA was subjected to injury repair, end-repair, ligated to SMRT dumbbell-type linkers, unligated linker sequences at both ends of cDNA were removed, primers were added, and DNA ...
All the candidate elements in 1p22 locus were cloned using the BAC RP11-109C4 as template and primers are documented in Supplementary Table 9. Products were cloned into pENTR/D-TOPO plasmid (Life Technologies, Carlsbad, CA, USA) for Sanger Sequencing validation. After comparing the gen- ...
5d). Using a two-step selection strategy, we first knocked in the desired allele in a template containing a neomycin-resistance (neo) cassette, and then removed the neo cassette, leaving behind a single loxP site of 34 base pairs (Supplementary Fig. 5a–d). As for HDR in E2, although ...