FloodLAMP Test Training Video Course
Video 07 - Adding Samples to Reaction

Shows the process of adding samples to strip tubes, including organization and labeling, careful handling of controls and samples to prevent cross-contamination, and the importance of following protocol steps for preparing and sealing the tubes for amplification, with additional emphasis on glove changes.


Test Trainer  [0:00](https://vimeo.com/668227040/c1dd7a8cac?ts=0)
Okay, so we're ready to add the samples and you can actually do this while they're cooling down. Our timer just went off so I'll do this part fast. But what we're going to do is we're going to finish filling out our form here. Number of reactions is going to be seven. Five samples plus the two controls. We will record the positive control ID in just a second. We're going to do two things here. We're going to, just to be careful, we're going to put these in number order. 43. Again, you can do this while they're cooling down, 32, 24, and 23. Okay, There we go. If you want to be extra careful, you can write the numbers down. Spot number three, four, five, six, seven. This is actually when you're Using a larger number of tubes in our lookup rack, this is a little more straightforward. Here with small numbers, I kind of like to have the backup. 23, 24, 32, 43, and 115. Okay so we're going to set up the tips to align with reactions. Should already be set up. I'm just going to use the leftmost column there. Okay, and now we're going to do something important and this is, should be in our protocol here. We already had done that. We'll go through that again. Here we go. This one right here. Strip 8 tube labeled on the top of each strip. So notice this guy isn't labeled, all nice and pink, so we're just going to put a one right there. Okay. Now, I'm going to get a piece of foil. I should keep this shut. A piece of foil for our caps. I like to try and save the caps. That way I don't get a mismatch between tubes and caps. If you do foul the caps or break it or have some problem, then don't worry about it, just get new ones. And here's a little technique tip, especially when these are colder. You do want to wait till it warms up that five minutes, but you kind of twist the first one off. And you're trying to make sure you don't touch the inside as you pull it off. So if your glove is not behaving, then try to fix it. Okay, and what I do here is I grab the tip and I'm pressing on the one behind it. We're not worried about cross-contamination here because there's no samples in here. So let's put that there, keep it nice and clean. Okay, we're going to do the negative control first. That is spot number two. Go down and get that. You always want to make sure you see the liquid in there. Go 12345. Blow out. Put These over here. Oh, don't try not to do that. There we go. Okay, we're gonna keep these guys covered while we work. Okay, get this out of the way, get our samples here ready to go, and we're ready to add them. Just going to put one at a time. Trying to hold the sample tubes over here, so hold them over here. Now these aren't activated, but you still don't want the chance for cross-contamination. So you've got to focus on counting to eight. Here's number three. There we go. Now you can look at the tip to see where you're at. Number four. There, I see it in the tip. Number four. If you lose track of where you are, just look right back over at the tips. That's why we align the tips, or set up the tips, we call it. There we go, number five. Number five. You'll get used to the positions here. If you get lost, look over there. Okay, and one more, six. If you get lost look over there, yeah okay and one more number seven. We're just going to leave that eighth one blank. Okay. Here we go. Keep these guys covered. So if we look at our list here, set up the tips, add the two microliters. You know, it doesn't say to add the positive control, it doesn't say to add the negative control because this is a short version. The long version has that in there. The positive control ID, actually had written down, it's TPC underscore. There we go. And I put quotes here for refer to the 1X. Okay. Got the positive control in the freezer here. So I'm going to be careful with this. I'll try to only touch the tube with my left hand. So what I do here is warm it up for about 30 seconds. So this is SARS-2 RNA, stabilized in total human RNA. This is the most fragile component of the test, meaning if you let it sit out at room temperature for very long, it degrades really fast and it won't work. Okay, so there we go. We're going to go back and get our first tip. We're gonna pipe it up and down to mix it quick. Oh, gotta make sure. That was something, I haven't had it a little too fast. You know, sometimes when you don't get your tip seated properly, it doesn't work. There, that worked. Doesn't work. There, that worked. Okay. And we got this. Okay. Shut this. Put this guy back in his little baggie. Back in the freezer. Okay, and a very important step here, which is to change gloves. If you just touch with your left hand and you're careful, you can just change one glove, but you have to change gloves. It's just a precaution to avoid any positive control cross contamination. So now we have our conveniently pre-loaded strip tubes here. When you grab these, be careful not to touch the inside. Okay, so press down there. There we go. Now, this critical part is you've got to make sure you hear that little pop. You've got to make sure that you're all pressed all the way down. Okay, and when you pull it out, you wanna look, make sure it's nice and flat. Okay, these guys are ready to go over here on the amp. All right, gotta put them in. Now, since I touched the amp area with my right hand, I'm going to change that glove and make sure I've got a timer set. I should have a separate timer over there, which I don't have right now. So let's just stop it a few seconds early since I started a few seconds late. Now I'm gonna show you, show you filling out the form when this comes off. All right.