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(DOC 281 KB) 12284_2014_5_MOESM3_ESM.doc Additional file 3: Figure S2.: Description of the sgRNA Cas9 binary vector and the expression of Cas9.

The information entropy for a system with a probability distribution P(x i ) for two classes information entropy [ 49] is defined as (3) H x = − ∑ x P (x i ) log ⁡ 2 P (x i )   (i = 0,1 ), where P(x i ) represents the occurrence probability of number i in the aforementioned 1024-bit binary vector and the information entropy H x is a measure value of the information amount.

ORFs were than cloned as BamHI- PstI fragments into the shuttle vector pBJ36-35 S. Cassettes containing the 35 S promoter, the FAF ORF and the ocs terminator were excised from the respective plasmids using NotI, ligated into the pMLBART binary vector and transformed into Col-0 wild-type plants by floral dipping [ 41].

We performed hierarchical clustering on each dimension of the matrix by using the Tanimoto coefficient (Tc) between binary vectors and then selected the major part in resultant clusters of each dimension for detailed analysis (Fig. 3 and Methods).

These are the binary vectors and can be maintained in both E. coli and A. tumefaciens.

Two variants are the super binary vector strategy and the twin T-DNAs strategy.

Amplified gene flanks are then mixed with specifically designed marker and binary vectors and treated with BP clonase, generating the deletion construct in a single cloning step.

Among the factors involved included the genotype of plants, types and age of tissues used, the Agrobacterium strains and binary vectors used, and the various tissue culture conditions [ 6].

Transgenic apricot and tobacco lines transformed with the binary vector pBin19-35SGusintron, and which carry the nptII gene, were obtained and their DNAs were serially diluted to obtain a standard curve for both the endogenous β- actin and nptII genes.

EHA105 containing the disarmed Ti plasmid and the binary vector pCAMBIA1301 containing the GUS gene with castor bean catalase intron, the kanamycin phosphotransferase II (ntpII) gene and hygromycin phosphotransferase (hpt) gene, conferring resistance to kanamycin and hygromycin respectively.

For selection of transgenic plants, HptII was isolated from the binary vector pCAMBIA1300 and inserted into the pZD264 vector using the PmeI (GTTTAAAC) cloning site, which resulted in a vector of p1500 for rice transformation.