All the sample DNAs from the L. vannamei, C. hongkongensis, organisms in pond water were amplified in duplicate PCRs at a 25 μL reaction volume, containing 10 μL PlatinumTM Hot Start PCR Master Mix (2X) (13000014, Thermo Fisher Scientific, Lithuania), 1 μL (each) primer 1391f and primer EukBr (10 μM), 1 μL template DNA, 8–12 μL nuclease-free water, and 0–4 μL blocking primer (10 μM). The PCR thermal cycling conditions were as follows: 3 min at 94°C; 35 cycles of 30 s at 94°C, 30 s at 55°C, 1 min at 72°C; and finally 10 min at 72°C. The PCR products were checked by electrophoresis on a 1.5% agarose gel stained with DNAGREEN (UV) (TIANDZ). In the electrophoresis results, the decrease in intensity of the band compares to the no-blocking, reflects the inhibitory effect of the blocking primer on the host DNA. The less intense the band, the better the blocking effect.
Dnagreen uv
Manufactured by Tiandz
Sourced in China
The DNAGREEN (UV) is a UV transilluminator designed for visualizing and analyzing nucleic acids, such as DNA and RNA. It utilizes UV light to excite fluorescent dyes that bind to the nucleic acids, allowing for their detection and documentation. The DNAGREEN (UV) provides a reliable method for viewing and imaging nucleic acid samples in a variety of laboratory applications.
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Lab products found in correlation
2 protocols using dnagreen uv
1
Blocking PCR Optimization for Environmental DNA
2
ISSR-PCR Amplification Protocol
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A total of 100 ISSR primers (University of British Columbia, Vancouver, BC, Canada) were tested with nine (UBC #808, #824, #827, #836, #841, #842, #847, #857, and #873) showing high reproducibility and accuracy selected for PCR amplification. The PCR reaction was performed as described in literature with a minor modification [36] (link). The optimal reaction condition of ISSR-PCR amplification contained 2.5 µL template DNA, 2 µL 10× PCR buffer, 1.5 µL dNTPs, 0.2 µL Taq polymerase, 0.5 µL primer, 13.3 µL ddH 2 O to make the total volume of 20 µL.
The cycling conditions were as follows: 4 min at 94 • C for an initiation step, followed by 40 cycles of 30 s at 94 • C, 30 s at a primer-appropriate temperature, and 2 min at 72 • C, and finalized at a final cycle of 10 min at 72 • C. PCR products were detected by electrophoresis on a 2.0% agarose gel stained with 0.1 µL/mL of DNAgreen (UV) (Tiandz, China) and run in 1× TAE buffer at 100 V for 40 min. PCR products were photographed by the Bio-image System (BioRad, Germany). Each PCR reaction was repeated three times.
The cycling conditions were as follows: 4 min at 94 • C for an initiation step, followed by 40 cycles of 30 s at 94 • C, 30 s at a primer-appropriate temperature, and 2 min at 72 • C, and finalized at a final cycle of 10 min at 72 • C. PCR products were detected by electrophoresis on a 2.0% agarose gel stained with 0.1 µL/mL of DNAgreen (UV) (Tiandz, China) and run in 1× TAE buffer at 100 V for 40 min. PCR products were photographed by the Bio-image System (BioRad, Germany). Each PCR reaction was repeated three times.
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