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Real-time PCR Mixture Preparation and Protocol
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Real-time PCR Mixture Preparation and Protocol

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Questions and Answers

Kastamonu garlic clone is suitable for summer consumption

False

Tunceli garlic is common in multiple provinces of Turkey

False

The garlic production areas and seedling production areas are separated in Turkey

False

Nematode is one of the most important diseases and pests affecting garlic production in Turkey

<p>True</p> Signup and view all the answers

The white rot disease does not affect garlic production in Turkey

<p>False</p> Signup and view all the answers

The most common viruses affecting garlic production in Turkey are OYDV and LYSV

<p>True</p> Signup and view all the answers

The viruses OYDV and LYSV together result in a yield loss of up to 50% in garlic production in Turkey

<p>False</p> Signup and view all the answers

Real-time PCR method is less sensitive and specific than other techniques for identifying plant viruses.

<p>False</p> Signup and view all the answers

The meristem culture technique is widely used for obtaining virus-free plants in garlic production.

<p>True</p> Signup and view all the answers

Extracting the meristem regions of the plant is easy and requires minimal expertise.

<p>False</p> Signup and view all the answers

Nucleic acid production capacity in meristematic tissue during cell division is used for the formation of virus particles.

<p>False</p> Signup and view all the answers

The study evaluated the micropropagation capacity of A. tuncelianum, an endemic species of Turkey, and A. sativum, the Kastamonu garlic clone.

<p>True</p> Signup and view all the answers

Transportation of viruses to the meristem region of the plant is prevented due to the lack of transport system in meristem.

<p>True</p> Signup and view all the answers

Real-time PCR method was found to be more sensitive than other methods in the diagnosis of potato virus Y (PVY).

<p>True</p> Signup and view all the answers

Two different nutrient media were tested in terms of micropropagation success, with Medium 2 found to be more successful than Medium 1.

<p>True</p> Signup and view all the answers

There is an effective chemical application available against virus control in garlic production in Turkey.

<p>False</p> Signup and view all the answers

Meristem culture technique is the most common method for obtaining virus-free plants

<p>True</p> Signup and view all the answers

Shoot tip culture technique is sufficient for all cases of virus-free plant propagation

<p>False</p> Signup and view all the answers

Real-time PCR assay can provide quantitative assessment of viruses in plants with high efficiency and very low chances of infection

<p>True</p> Signup and view all the answers

The study was conducted at the University of Cukurova Turkey and Adana Veterinary Control Institute between 2009 and 2010

<p>True</p> Signup and view all the answers

Allium sativum (Kastamonu garlic clone) and Allium tuncelianum garlic species infected with viruses were used as plant material

<p>True</p> Signup and view all the answers

Sixty in vitro plants were used for real-time PCR studies after 6 weeks of micropropagation

<p>True</p> Signup and view all the answers

The qualitative detection program was not used for evaluation in the Roche LightCycler 2.0 Real-time PCR

<p>False</p> Signup and view all the answers

Real-time PCR was carried out using 600s at 95°C for denaturation, 45 cycles of 10s at 95°C, 30s at 60°C, 2m at 72°C, and 30m at 40°C for cooling.

<p>True</p> Signup and view all the answers

The primers used for OYDV and LYSV viruses were not registered in GenBank and their sequences were not specified.

<p>False</p> Signup and view all the answers

Ninety meristems and ninety shoot tips were used in the experiment for A. tuncelianum.

<p>False</p> Signup and view all the answers

Meristem cultures resulted in 84 healthy plants, 1 unhealthy plant, and 5 infected plants for A. tuncelianum.

<p>True</p> Signup and view all the answers

The plantlets obtained from meristem cultures showed better micropropagation results in Medium 2 compared to Medium 1 for A. tuncelianum.

<p>True</p> Signup and view all the answers

In the first micropropagation of A. sativum, 89 meristems and 87 shoot tips were used.

<p>True</p> Signup and view all the answers

The study concluded that shoot tip culture was more effective than meristem culture in obtaining virus-free plantlets.

<p>False</p> Signup and view all the answers

Study Notes

  • A mixture was prepared for real-time PCR detection of OYDV and LYSV viruses in A. tuncelianum using SYBER Green. The mixture contained isolated DNA, SYBER Green master, primers, Mg, and water.
  • The real-time PCR was carried out using 600s at 95°C for denaturation, 45 cycles of 10s at 95°C, 30s at 60°C, 2m at 72°C, and 30m at 40°C for cooling.
  • The primers used for OYDV and LYSV viruses were registered in GenBank and their sequences were specified.
  • Ninety meristems and ninety shoot tips were used in the experiment for A. tuncelianum. Meristems took 8 weeks to grow into plants, while shoot tips took 3-4 weeks.
  • Meristem cultures resulted in 84 healthy plants, 1 unhealthy plant, and 5 infected plants. Shoot tip cultures resulted in 80 healthy plants, 1 unhealthy plant, and 9 infected plants.
  • The plantlets obtained from meristem cultures showed better micropropagation results in Medium 2 compared to Medium 1. In the first micropropagation, 14.1 shoots/plant and 4.63 shoots/plant were obtained from Medium 2 and Medium 1, respectively.
  • In the first micropropagation of A. sativum, 89 meristems and 87 shoot tips were used. Meristem cultures resulted in 89 healthy plants and 1 unhealthy plant. Shoot tip cultures resulted in 87 healthy plants and 3 infected plants.
  • The micropropagation results showed that Medium 2 was more successful than Medium 1 in both garlic species.
  • Sixty garlic plants were tested for OYDV and LYSV viruses using real-time PCR. No viruses were detected in meristem cultures of both garlic species. Viruses were detected in shoot tip cultures of both garlic species, with OYDV detected in 80% of tested plants and LYSV in 67% of tested plants for A. tuncelianum, and 73% and 87% for A. sativum, respectively.
  • The study concluded that meristem culture was more effective than shoot tip culture in obtaining virus-free plantlets.

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Description

Learn about the preparation of a real-time PCR mixture for DNA detection using SYBER Green master and the protocol for the detection process. This quiz covers the components of the mixture, the PCR protocol, and the use of SYBER Green for DNA detection.

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