ITS2Internal Transcribed Spacer Region 2 (DNA)
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A combination of ITS-1 and ITS2 sequences has been used to identify the C.
Several methods for specific fungal species identification are based on ITS2 ribosomal region variability, which is flanked by conserved sequences of rDNA (Fujita et al., 2001; Ferrer et al., 2001; Trost et al., 2004).
Reactions for 12S, ITS2 and 28S were run at 95[degrees]C for 5 min; then 35 cycles of 95[degrees]C for 30 s, the annealing temperature for 30 s, and 72[degrees]C for 60 s; with a final extension at 72[degrees]C for 10 min.
Phylogenetic inference from ITS2 is typically problematic due to large interspecific differences that make alignment of this region difficult and somewhat ambiguous.
Polymerase chain reaction (PCR) was performed using the following Symbiodinium-speciuc ITS2 primers: forward primer (ITS2infor) 5'-GAATTGCAGA AC TCCGTG-3' and reverse primer (ITS2CLAMP) 5'-CGCCC GCCGC GCCCCGCGCC CGTCCCGCCG CCCCCGCCC GGGATCCATA TGCTTAAGTT CAGCGGGT-3'.
Bogado, "Solution of the point kinetics equations with temperature feedback by ITS2 method," Prog.
The 25S and 18S probes comprise the entire rDNA gene sequences (3.2 and 1.8 Kbp, respectively), while 5.8S/ITS probe (0.7 Kbp) involves the 5.8S gene and its flanking ITS1 and ITS2. The IGS4 probe (0.5 Kbp) consists of structural regions (SR) III-VI of the rDNA B-type IGS (2 Kbp) of Capsicum and includes the putative transcription initiation site motive (TIS) and diverse postulated regulatory elements widely conserved in Solanaceae.
Methods: We assessed the validity and utility of standard DNA barcoding in the quality control of raw materials, extracts and finished products in 61 samples using four universal barcodes (matK, rbcL, trnH-psbA and ITS2).
Sequences of the universal primers for evaluating DNA barcodes, including those for ITS1, ITS2 (ITS1 + ITS2, herein named as ITS region) and rpoCl, and general PCR reaction conditions, were obtained from previous studies (Tokuoka & Tobe, 2006; Chen et al., 2010; Sharma, Folch, Cardoso-Taketa, Lorence, & Villarreal, 2012).
tropicalis and W saturnus, containing 18S rDNA (partial), ITS1 (complete), 5.8S rDNA (complete), ITS2 (complete), and 28S rDNA (partial).
In order to resolve the difficulties in amplification and sequencing of the ITS region, they suggested the use of the short ITS2 region as a backup because of the presence of it conserve sequences [224].