Exact(4)
Specific RT primers for the miR-29b sequence, guide strand (5′-UAGCACCAUUUGAAAUCAGUGUU-3′ Qiagen), or U6 RNA were used.
The crRNA consists of a conserved repeat sequence and a variable spacer sequence (guide) complementary to the target gene sequence (Brouns et al. 2008; Hale et al. 2009).
The expression vectors for the gRNAs were generated by ordering a 455 bp DNA fragment (composed of U6 promoter, target sequence, guide RNA scaffold and termination signal) as gBlocks from IDT (Leuven, Belgium) and then cloning them in pUC19 plasmid.
RNA oligonucleotides for FUS (sequences below) and a scrambled control sequence (guide: 5′-AAUUCUCCGAACGUGUCACGU-3′ passenger: 5′-GUGACACGUUCGGAGAAUCUU-3′) were purchased through Sigma-Aldrich and 10 μM annealed stocks were prepared by combining guide and passenger strands in the following buffer: 100 mM potassium acetate, 30mM HEPES-KOH pH 7.4, 2 mM magnesium acetate.
Similar(56)
An ingenious new calibration sequence guides you through adjusting your set for the best picture.
The engineered DNA binding protein or RNA sequence guides the nuclease to a specific target sequence in the genome to induce a double strand break.
When describing expression patterns, a gene or an "ID" (shorthand for "identifier") therefore refers to a predicted transcript whose sequence guided the design of a Probe Set on the Lotus Affymetrix array.
Wards will adopt HEADS-UP in a pragmatic sequence, guided by local clinical enthusiasm.
These RNAs, ~22 nucleotides in size, act as sequence guides that direct Argonaute protein complexes to mRNAs, where they decrease protein synthesis through translational repression or mRNA degradation and thereby influence many basic cellular processes and diseases [ 52- 58].
Also, the mechanism by which DNA sequences guide nucleosomes positions is different between S. pombe and S. cerevisiae [ 18].
For both organelles, signal sequences guide the protein through the outer and, where appropriate, inner membrane via protein translocases ("translocons").
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