Journal: Scientific Reports

Article Title: Targeted mutagenesis in soybean using the CRISPR-Cas9 system

doi: 10.1038/srep10342

Figure Lengend Snippet: Construction of binary vectors for genome editing in soybean. Cas9 fused with a single nuclear localization signal (NLS) is expressed with a Cauliflower mosaic virus 35s (CaMV 35s) promoter. Synthetic guide RNA (sgRNA) is derived using U6 promoters. ( a ) Arabidopsis thaliana U6-26 promoter ( b ) Glycine max U6-10 promoter. Sequences containing two Bsa I sites are located between the U6 promoter and the sgRNA scaffold. These sequences can be easily replaced with a gene-specific sgRNA seed. LB: left border; RB: right border.

Article Snippet: Arabidopsis U6-26 and soybean U6-10 promoters with sgRNA were synthesized (Genscript, Nanjing, China) ( and ) and cloned into pUC57-Kan vectors to generate pUC57-AtU6-26-sgRNA and pUC57-GmU6-10-sgRNA plasmids, respectively.

Techniques: Virus, Derivative Assay

Journal: Nucleic Acids Research

Article Title: Short tandem repeat stutter model inferred from direct measurement of in vitro stutter noise

doi: 10.1093/nar/gky1318

Figure Lengend Snippet: The synthetic STR experiment summary. ( A ) Schematic description of the synthetic library. In each plasmid, a different synthetic STR construct was designed, synthesized and clone-sequenced for various STR types and length. The STR was designed within a context of an Illumina Truseq-HT dual index library to enable for nested PCR amplification at two time points (T 2 - amplification using outer primers only, T 3 -amplification using inner primers followed amplification by outer primers). The library is flanked by BsrDI restriction sites to enable direct sequencing of the STR library without amplification (T 1 ). Internal barcode (yellow triangle) is a short sequence, unique to each STR length to detect for cross-contamination. See text and methods for elaboration and for the designed constructs. ( B ) AC STRs repeat-number histograms, as were interpreted from sequencing results (T 1 , T 2 and T 3 ), compared to their expected length, T 0 (designed sequence). ( C ) Sequencing analysis results of each STR type, repeat-number and time point described as the percentage of the original (designed) signal from all the reads. Dashed line at the 5% marks the lower threshold of analysis: data points below the mark were deemed too noisy and were excluded from downstream analysis.

Article Snippet: STR plasmid design: Sequence verified cloned plasmids containing synthetic STRs of different types and sizes ( ) were ordered from either IDT or GenScript (pIDT-kan and modified puc57-Kan vectors, respectively).

Techniques: Plasmid Preparation, Construct, Synthesized, Nested PCR, Amplification, Sequencing