Genomic DNA was isolated from 16 days old culture in log phase using the Himedia Ultrasensitive Spin Purification Kit (MB505) following the instructions of the manufacturer, except for the increase of incubation time for the lysis solutions Al and C1, which were set to 60 and 20 min, respectively. DNA fragments within the following genes were amplified using the oligonucleotide primers and PCR programs listed in Table S1 . PCR products were checked by electrophoresis on 1% agarose gels (SeaPlaque® GTG®, Cambrex Corporation), using standard protocols. The products were purified directly using the Geneclean® Turbo kit (Qbiogene, MP Biomedicals) and sequenced using the BigDye® Terminator v3.1 cycle sequencing kit (Applied Biosystems, Life Technologies). The partial sequences were compared with the ones available in the NCBI database using BLASTN. The BLAST X tool (blast.ncbi.nlm.nih.gov/Blast.cgi ) was used for determination of the nos, mcy G and mcy D genes similarity. The sequences were annotated for the coding regions by the NCBI ORF Finder at NCBI (http://www.ncbi.nlm.nih.gov/gorf/gorf.html ) and the ExPASY proteomics server (http://www.expasy.org/tools/dna.html ).
Ultrasensitive spin purification kit
Manufactured by HiMedia
The Ultrasensitive Spin Purification Kit is a laboratory equipment designed for the purification of DNA and RNA samples. The kit utilizes a spin column-based method to efficiently extract and concentrate target molecules from complex biological samples.
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Lab products found in correlation
2 protocols using ultrasensitive spin purification kit
1
Genomic DNA Isolation and Sequence Analysis
2
Isolation and Amplification of rbcl Gene
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The DNA was isolated from 12-day-old cultures using Himedia Ultrasensitive Spin Purification Kit (MB505) with some modifications (Singh et al. 2013 ). The DNA was stored at -20 °C. The rbcl gene was amplified using the primer pairs rbclf (5′-GACTTCACCAAAGAYGACGAAAACAT-3′) and rbclr (5′-GAACTCGAACTTRATYTCTTTCCA-3′) (Halinen et al. 2007 ). The amplification reaction was carried out at an initial denaturation of DNA at 94 °C for 5 min, followed by 30 cycles of denaturation at 92 °C for 1 min, annealing at 55 °C for 1 min, and extension at 72 °C for 2 min. The final extension was done at 72 °C for 6 min, followed by incubation at 4 °C for 20 min. The appropriate sized bands of 700 bp were cut using sharp aseptic blades and sent for sequencing.
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