Text Transcript with Description of Visuals
| Audio | Video |
|---|---|
| [Music] Hello and welcome to the total target for sprout damage protocol. This test measures alpha-Amylase enzyme activity and provides an estimated falling number in a milled wheat sample. You may perform this protocol on up to 6 samples at a time. This video shows a batch of 3 samples. | Text reads, Total Target for Sprout Damage Protocol, instructional video. |
| First, gather your materials. | Text on screen reads, Gather Materials |
| Your test kit contains pipette tips, 5 milliliter reaction tubes, DB6 stop buffer, substrate tablets, and test strips. Kit items must be left at room temperature for at least 30 minutes prior to testing. | A video shows a text kit with text, Envirologix, on the side. The Narrator explains the items. |
| You will also need a QuickScan II machine, mini centrifuge, Vortexer, dry bath preheated to 70 degrees Celsius, scissors, pipette, tweezers, and a permanent marker. | Text reads, Equipment. Images appear of the items needed. |
| [Music] | Text on screen reads, Disposable Materials |
| Disposable materials required are 1 and a half millileter microfuge tubes, deionized water, and a way to accurately measure and dispense the water, and a disposable container with water-tight lid for each sample. | Images appear of the items listed. |
| Set up your workstation in a way that is comfortable and provides easy access to each piece of equipment. | The camera pans across a workstation showing all of the previously mentioned equipment arranged neatly. |
| Set up the QuickScan terminal. Turn on your QuickScan II machine and open the QuickScan app. | Text on screen reads, Setup: QuickScan Terminal |
| If the machine does not calibrate automatically, select the calibrate button. | A camera view of the QuickScan screen is shown. EnviroLogix QuickScan software is shown on the screen. In the top right, a circle has the text, read test inside it. Across the bottom of the screen of the Quick scan 2 software, there are several links. Help. Calibrate. Clean Test. Data Log. Set Up and Exit. A square appears around, calibrate. |
| Perform a clean test by inserting a white sheet of card stock into the clips located at the back of the tray. Press clean test. | A tray is slid open on the Quick scan 2 and gloved hands insert a piece of card. The Narrator explains. |
| The results should indicate zero bad clusters. | A close-up view of the screen shows the text, 0 bad clusters. A red rectangle appears around the text. |
| Next, place the check cone face up in the carrier tray with the bar code area closest to the front. | A blue-gloved hand places a small card with several cutouts into the tray of the machine and closes the tray. |
| Press read test on the QuickScan home screen. | A blue-gloved hand taps the on-screen button that has text that says, Read Test. |
| All results should be reported as okay. | A view of the screen shows the test results. In the results column all six entries read okay. |
| [Music] | Text on screen reads, Running the Test. |
| Label your sample, microfuge and reaction tubes according to the number of samples to be run per batch. For example, our tubes are marked 1 through 3 for each step. | Numbers 1, 2, and 3 are written on the lids of sample tubes. As the video continues, the Narrator explains. |
| Gather your samples for this batch. | A camera view of the workstation shows all of the equipment arranged neatly on a bench top. A person wearing a white lab coat and blue gloves stands at the bench. In the upper-left corner of the frame, a close-up view of three labeled tubes is superimposed over the view of the bench top. |
| Each sample tube should contain 5 grams of milled wheat. Add 25 milliliters of deionized water into each sample tube, add the lids and tighten. [Music] | The person removes each of the three lids from the three tubes and adds a measured amount of liquid to each tube with a syringe. Later, a fast-forward icon appears in the lower left corner of the frame. Below this icon, the text 2x appears, indicating that the video has been sped up to 2x speed. The person replaces all three lids on the tubes. |
| [ Music ] Turn the tubes upside down and tap onto your work surface, until the flour releases. | The person follows the protocol as described. |
| [ Music ] Set a timer for 30 seconds and shake to incorporate the flour and water. | The person follows the protocol as described. |
| [ Music ] Return the tubes to their rack. Being sure to keep them in numerical order. | The person follows the protocol as described. |
| [ Music ] Transfer an equal volume of each sample into a labeled 1 and a half milliliter microfuge tube. The tubes can be three quarters to fully filled, as long as the volume is consistent between the tubes. Make sure to change out your pipette tip or your disposable pipette between samples to avoid contamination. | The person follows the protocol as described. |
| [ Music ] Load samples into a mini centrifuge, making sure that the samples are evenly spaced for balance. If you are running an odd number of samples, you may use a tube filled with water to achieve a balanced centrifuge. Run the centrifuge at 6,000 RPM for 30 seconds. | The person follows the protocol as described. |
| [ Music ] Remove the samples. As you can see here, the solid material has gathered at the bottom of the tube, leaving a clarified liquid at the top. | The person follows the protocol as described. A close-up of the tube is shown with the solids and liquids separated. |
| [ Music ] Use tweezers to add 1 substrate tablet to each reaction tube. Make sure not to touch the tablet, as it can interfere with test results. | The person follows the protocol as described. |
| [ Music ] Pipette 300 microliters of the clarified liquid from each sample into its respective reaction tube. If enzymes are present in this liquid, they will start feeding on the substrate tablet. Again, make sure to switch out your pipette tip between samples. | The person follows the protocol as described. |
| [ Music ] Vortex for 15 seconds to break down the substrate tablet and incorporate the mixture. | The person follows the protocol as described. |
| [ Music ] Place reaction tubes in a dry bath preheated to 70 degrees Celsius and immediately start a timer for 90 seconds. The heat from the dry bath speeds up the reaction, so that we can get a faster result. | The person follows the protocol as described. |
| [ Music ] At the 90-second mark, quickly add 300 microliters of DB6 stop buffer to each reaction tube, while the tubes remain in the dry bath. | The person follows the protocol as described. |
| [ Music ] Remove the reaction tubes from the dry bath and pulse 3 or 4 times on the Vortexer to mix. Return the reaction tubes to their rack and allow them to settle for 30 seconds at room temperature. Make sure that you've put them back in the correct order. During this time, prepare 1 test strip for each sample, if you have not already done so. The bar code end of the strip may be handled, but the end with the arrows pointing downward is the reactive pad and should not be touched. | The person follows the protocol as described. |
| [ Music ] If desired, you may keep the strips in a set of empty tubes for easier transfer. You may also place small marks or numbers on the back of the test strips, to ensure they are kept in the correct order. | The person follows the protocol as described. |
| [ Music ] Place test strips into the reaction tubes and set a timer for 3 minutes. [ Music ] At exactly 3 minutes, remove strips and cut off the reactive pads. | The person follows the protocol as described. |
| [ Music ] Load the strips into the QuickScan II reader, being very careful to keep them in the correct order. Strips are loaded from right to left. Close the tray. Press read test and follow the prompts on the screen to view your results. [ Music ] | The person follows the protocol as described. |
| [ Music ] | The results of the test appear on the screen. |
| [ Music ] | Text on screen reads, Thank you for watching! |
| [ Music ] | Text on screen reads, This video was made possible by funding from the Foundation for Food & Agricultural Research (FFAR) Grant number, 22 dash 000210. Matching FFAR funding was also provided by Envirologix, High Line Grain Growers, The McGregor Company, The Washington Grain Commission, The Wheat Marketing Center, and Washington State University. Logos appear for the above institutions. |