Mr. Yamauchi, you had mentioned that the next tube in order was the cloth control. Can you explain to the jury what the function of the cloth control is in the PCR DQ-Alpha processing?
It acts as a negative control in the analytical process for that particular step. From that point on, it would show us whether something funny happened or not.
When you say "Show us whether something funny happened," can you give us an idea what you mean by that?
Well, it's kind of inherent in the way I described that. By negative control, we expect it to come up negative. But if it comes up and it shows that there's some activity or something there, then it's quite possible some of our solutions at that point could have been contaminated. And this would be a way that we can double-check and know later on if something was astray or different that--at this point, a contaminant was present.
That control? Like I was saying before, generally speaking, we expect it to be negative. But if some activity comes up throughout the whole process where we see something on a strip, then we know that perhaps something happened at that stage and it would help us and aid us in troubleshooting.
And what about the next control in order after the cloth control on this first set of DQ-Alpha typing? What was that after the cloth control?
Your Honor, at this point, I would like to have marked as People's 281 for identification a photo display board, the LAPD PCR typing board.
Okay. Mr. Yamauchi, you've generally described the extraction procedure that you utilized in this case. And referring to People's 281 for identification, could you step down and, using the pointer, could you please describe what's shown in the photographs starting at the top horizontal strip that we've boxed in? It's entitled "Piper tech extraction." Now, the jury has heard what piper tech is, but you mentioned that the extraction is in the serology room?
Okay. And in the left-hand photo under "Piper tech extraction," could you explain what you are doing in that photograph?
Okay. At this point right here, the first step is to add one unknown of sterile water. And that's to get out some extraneous components that may or may not be a problem. And one case in point would be the particular group that's labeled "Blood, red." There's no inhibitor of PCR process. So at this point here, by adding the water, it puts that group into the inclusion and it can later be separated from the DNA that were actually after.
Mr. Yamauchi, could you try to keep your voice up because you've got your back to the Court reporter. I know it's important that we all get to hear you, okay? Okay. What's in the next photograph, the middle photograph, under pipe tech extraction?
And is that the same tube that you started out by putting the cuttings that you made over in the evidence processing room there or is that a different one?
Okay. And then over in the right-hand side, what is that that you're doing and what is that machine?
Oh, okay. After the water is placed into the tubes and allowed to soak for a--about 20 minutes or so, it's then put into the--what's called centrifuge--microcentrifuge--microcentrifuge tubes. This is what I'm doing. Goes into that instrument and spins it down at a high volume. All the solid components or heavy components like DNA, for example, are going to be forced towards the bottom while the other components that are in solution will just remain in solution.
Okay. Now, what do you do with these tubes that are--after they've been centrifuged at the end of the process?
At this point, there's another step called--it's kind of a mixture of chemical called chelex and it acts in a specific function in the extraction process which--and this is kind of weird because they're not really sure what its actual action is, but they do know that it does aid in taking out a component that could help in the degradation of DNA. So to a certain extent, it acts as kind of preservative.
Now, are there different forms of extraction processes that one can do in order to do the PCR DQ-Alpha typing?
Chelex is the one we use at our lab or the one that I use in this case. There's also what's known as a pheno chloroform extraction.
Okay. Now, are you familiar with the amplitype user guide for PCR DQ-Alpha typing that's provided by the manufacturer?
Well, generally speaking, that's the procedure that we use or I use personally most often, and that would be one reason. The other reason would be that it actually has lesser steps and it is faster.
Okay. And do you process the evidence--or strike that. When you perform the extraction process, is it your practice to never open more than one tube at a time?
Mr. Yamauchi, how many tubes do you open at any given time during the extraction process?
Now, is--could you describe how long the chelex extraction process takes in comparison with the organic extraction process?
Well, I haven't done the organic one in a while, so I would have to reference that to get a time frame on it. But as far as the chelex is concerned for bloodstains, it's in an hour and a half about you can get the extraction completed.
How long does it usually take you to perform the chelex extraction process in other cases?
After they're put in a rack, see, the next step would be amplification. So that rack of tubes would have to go over to where we have our thermal cycler, and that's the machine we use to do the amplification. We have that setup in our laboratory over at Parker Center facility. So there's a space difference of about a couple miles maybe or so, hop in the car and drive over there.
Now, are you familiar with the provisions in the amplitype user guide with respect to the relationship of the amplification room to other work areas in the PCR DQ-Alpha process recommendations?
The recommendations, safety precautions, they want all these rooms basically separated off in space and especially the amplification room and extraction and examination areas.
To eliminate the chances of cross-contamination. Especially of interest at this point is, when you're dealing with the amplification area, there is a second type of contaminant called the--well, you've probably heard this already--PCR carry-over. It's where part of the PCR process is more likely to contamination because of its perfect size and sequence. It's more likely to be a harmful contaminant. That's why we take extra precautions to keep this area physically out of the way of the other areas where we do our extractions and our evidence analyzing.
How do you get from piper tech serology to Parker Center? How did you get there on June 14th in this case?
Well, the closed-off tubes are in the rack. And then one of those chem-wipes I described earlier, there's larger versions, I'll wrap that whole rack and everything up in the chem-wipe and transport it that way.
Mostly because when you're walking along around the police station, it makes the detectives and people over there kind of nervous if they see little test-tube things. So I just want to make sure it's covered up.
KEY QUOTECan you describe the facility or the rooms that you have over there? Are they normally staffed by anybody?
Well, I have a key and Erin Riley does and Harry Klan because we work in that area. I believe some supervisors do too.
Okay. Do you recall about what time it was--or strike that. Let's just talk generally. You get over to the--you got your key to the amplification area, Parker Center. Will you describe what's shown in the photograph--on the left-hand side, it's labeled, "Pre-amplification." What are you doing there?
This is a room that's separated from the amplification room. Amplification room is the one with the instrument thermocycler and this--in the pre-amplification area is where we set the tubes up, process them with the right chemicals to be taken over and placed into the machine.
And you've described the pre-amplification--is that the same room as the amplification takes place?
Okay. So the photo in the left-hand side shows what you've just described, and that's in one room, and then the next photo, it's labeled "Amplification," what's going on in that photograph?
That's where the actual amplification process takes place. That's where the thermal cycler machine is housed, in that room.
And at this point in the process, are there additional controls that are added to the controls that you've described in the extraction process?
Well, those would be set up over here, but yes, there are. For the amplification process, we have another positive and negative control.
Well, like I was saying, before you have the controls that we take at the scene, we've got controls at that extraction point. We also have another set of controls at this point where we're going to run the amplification just to give us more information again.
What's shown in the next photo on the right, the hibridization--the one that's labeled "Hybridization," what are you doing there?
That's to display the hybridization strips that we have, and those are the strips that contained the probes that we eventually get our results from.
Okay. At Parker Center, after we do the strips, we've got to photograph them, and that's the area over there at Parker Center that we do that.
Because the strips themselves, they tend to discolor. They won't be as good as that original photograph if you hold on to them. So we have no reason to keep them.
At this point, generally speaking, as you do the procedure in Los Angeles Police Department, you've got photographs of the strips, and the board has another line entitled "Piper tech electrophoresis." Can you describe what's in the left-hand photo, picture of those doors?
And is there some sort of security device that monitors people coming in and out of the instrument room?
Now, what do you need to take from the Parker Center at the end of the amplification and hybridization back to piper tech to take into the instrument room? What are you actually taking back there?
Have to take some of the amplified product from this--this reagent that's held in the thermal cycler, and that has to be brought back to piper tech to be analyzed in this product gel.
It's the kind to tell you whether or not the amplification process took place, and it can also give you more information as to a relative idea how much DNA was formed by the amplification although that's not real accurate.
A little while ago, you mentioned that it was important to avoid PCR product carry-over to keep the amplification separate from other areas.
No. The reason we have it in the instrument room is because it's separated physically from serology where we do our extractions.
Are you familiar with other laboratories where all of these functions are done within the same building, but in different rooms?
You wind up with electrophoresis gel where the product is moved through the electrical field and so it can be visualized and noted.
Now, this board doesn't actually reflect everything in the same position and location as it was when you performed your tests on June 14th and June 15th, does it?
--was changed. We were on a bench area that was kind of around the corner from that at the time that I actually analyzed evidence in the Simpson case. But since then, we've had to move our stuff over to the other corner.
Now, you've described the samples that you extract--that you sampled and that you extracted. Did you subject them to the PCR DQ-Alpha testing process in the manner that--in the general manner and sequence that is shown on the exhibit 281, the photo display board?
Okay. And if you can, I would like you to break down the process and ask about how long each step took, if that's possible to do. For example, how long did the chelex extraction process take you on June 14th for the samples that you've described to this jury?
Okay. And then from the extraction process, did you go right over to Parker Center after that?
Well, I don't know when I had my lunch that day, but that would have been the next step. Eventually, I would have had to drive over there.
KEY QUOTEAnd you process them, amplification, hybridization and photography, in the manner that's shown on the photo board 281 for identification?
Okay. And did you obtain PCR DQ-Alpha results on samples that are in the list of samples that you've previously described to the jury?
Mostly because when you're walking along around the police station, it makes the detectives and people over there kind of nervous if they see little test-tube things. So I just want to make sure it's covered up.
We throw the strips away after we're done. Because the strips themselves, they tend to discolor. They won't be as good as that original photograph if you hold on to them. So we have no reason to keep them.
He asked the same question. He always does that.
I don't know when I had my lunch that day, but that would have been the next step. Eventually, I would have had to drive over there.