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ImmunoStrip tests for the detection of Ralstonia solanacearum Catalog number: STX 33900 and ISK 33900
CONTENTS
ImmunoStrip
Item ImmunoStrip, blue label Sample bag containing BEB1 buffer* Instruction STX /0005 5 strips
not included
ImmunoStrip Kits
STX /0250 250 strips
not included
STX /0025 25 strips
not included
ISK /0005 5 strip 5 bags 1
ISK /0025 25strips 25 bags 1
ISK /0250 250 strips 250 bags 1
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*The Rs ImmunoStrip test must be used with BEB1 buffer.
YOU WILL NEED
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Scissors, a pen, and a knife or razor blade BEB1 sample extraction buffer, available as: o Mesh sample bag filled with BEB1 buffer (Agdia catalog no. ACC 00780) o BEB1 powder, 24.5 grams, makes 1 liter (Agdia catalog no. ACC 00066/025.4) 1.5 mlmicrotubes, pipette tips, and micropipette – when working with bacterial culture samples
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STORAGE
Keep the strips tightly sealed in the container with the desiccant at all times. Store container in the refrigerator (4°C) between uses. The sample buffer should also be refrigerated (4°C) when not in use.
BEFORE YOU BEGIN
Remove the kit from the refrigerator and allow the components towarm at room temperature (15-30°C or 60-85°F) for about 15 minutes. You will notice the cold BEB1 extraction buffer will contain a white precipitate or appear cloudy. The BEB1 will become clear after warming which signals that the test components are ready to use.
INTENDED USE
This test is a rapid means of screening crops for Ralstonia solanacearum (Rs). The Rs ImmunoStrip is intended for usewith plant samples exhibiting symptoms of Rs. The test can also be used to test bacterial culture samples. The Rs ImmunoStrip detects Rs to the species level and cannot differentiate race or biovar.
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ImmunoStrip for Rs
ImmunoStrip tests for the detection of Ralstonia solanacearum Catalog number: STX 33900 and ISK 33900
Sample Preparation
Plant samples: Samples should be takenfrom stems or petioles (Figure 1) of plants showing signs of infection (wilting). Samples are then ground in buffer at a 1:20 w/v ratio. This instruction describes the use of Agdia sample extract bags, though other means of extracting samples are acceptable. Sample extract bags contain 3 ml of BEB1 sample extract buffer that according to the ratio would require 0.15 g of tissue. When working withstems cut two cross section pieces at the first two internodes from the crown. The stem can be cut into smaller sections for easier grinding. Note: If you are using a knife or razor blade to cut samples, disinfect the cutting area and the knife or razor blade with alcohol between each sample.
Figure 1.
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Cut off the top of the sample extract bag, being careful not to spillthe buffer. Place the sample pieces between the mesh linings of the sample extract bag containing BEB1 extraction buffer. Rub the bag with a pen or blunt object to completely crush and mix the sample. Use only one sample per bag and be sure to label each bag.
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Bacterial culture samples: Bacterial culture samples can also be tested. Use a toothpick to remove a colony of bacteria from aculture plate. Stir into a tube containing 300 µl of BEB1 extract buffer. If using cell culture 6 broth, the cell suspension should be at a concentration of 10 cfu/ml or above. Add 50 µl of the cell suspension to a tube containing 250 µl of BEB1 sample extract buffer.
Grind 0.15 g of plant tissue in 3 ml of BEB1 sample extract buffer.
Instructions for use
Remove one strip from the packaging.When handling the strip, always grasp the top of the strip marked with the test name. Do not remove protective covering.
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Keeping the strip in a vertical position, insert the end of the strip marked “sample” into the extract. The strip should remain vertical during the test. Do not allow much more than 0.5 cm or ¼ inch of the end of the strip to be submerged in the extract. Be sure...
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