Lab 9 Immunoblot Report

I'm thinking about just letting this one be my dropped lab report. What the fuck is with asking us questions in the Data section? Where the fuck am I supposed to put it?

God I want to just get this semester done with

I just put my data questions in the data section and then did the rest in the questions section. The only one I am not sure on is question #7.

Question 7 is similar to the dilution question on the ELISA quiz, back page. I'm still playing with how exactly to do this one, too.

I used 0.5 ml of protein to 4.5 ml of buffer. 4 dilutions total. Does that sound right?

Does the standard curve have to be hand drawn? Cus i REALLY dont feel like doing it.....

It's just like the SDS-PAGE graph, so if you can do the semi log in Excel then I would say go for it, but I found it easier to just draw it.

this was a really long lab report - should have been worth double

my kalediscope colors dont match up to the appendix.... what should i do.....

What are you seeing?

Has anyone else figured out #7 yet? That is all I have left and I can print this stupid thing!

I'm sick of drawing best-fit lines. This will be my dropped lab grade, because I choose not to do it. Should we be expecting to be creating standard curves all the time with the types of jobs we occupy? Cause I might be in the wrong major. I LOVED Biology 3, and while I think a lot of our labs done are very cool, I don't like some types of analysis.

I guess beggars can't be choosers though in this type of economy. But godamn, I'm tired of getting reports back a month after I've completed them, and then being tested on them. I realize that Bradshaw is reasonable, but man do I wish I could just get done with this quarter.

1) kyle, calm down, it is oooook. breath! and honestly, in real life, you will probably have a computer program like excel or somethin do it for you. She wants us to do hand drawns so we know the mechanics behind it, ya know. but its ok.... you'll be fine.

2) the colors i see are.... no color but we have a dot there, so i assume its blue.... then i see magenta, then green.... then there is a space where there is like.... no color, but there is a dot.... but then the next color is clearly a purplish color then orange, then the rest ran down to the dye front which is kind of purple is so i think the red and blue ran together. So I dont understand... what should i call the middle portion that looks blank? I'm group 5 if you want to check out the pic.....

If you have a dot there, then you should have measured it. I asked her this because we do not have the bottom 2 bands of color on ours and did not have dots there either. She said to only use the ones with dots because we can't make up data that doesn't exist. Ours shows Blue, Magenta, Green, Violet and Orange in that order from top to bottom. Our protein shows up between the green and violet.

Sorry I didn't intend to bring anyone down to my level; I'm at a point where it is really hard to care about what constitutes an A, since none of our professors really line it out, and it seems like many put themselves > the class. (I.E. ignore me if you want a good grade.)

which group is your pic?

LWhitacre wrote:

I used 0.5 ml of protein to 4.5 ml of buffer. 4 dilutions total. Does that sound right?

Yes, this sounds correct.... same way i did it....

We are group 2

it looks llike the same thing as what i'm talking about. unless you didnt count that? i seee that the sample is in between the green and violet, but there is a dot there.... same as ours..... what color did you mark that as being? see what i mean?

Okay, so you set up 4 tubes. Start with 5 ml in the 1st, then have 4.5 ml of buffer in each tube. Then pipet .5 ml into each one. In the 4th tube you would have 5 ml at 1/10000. That sounds right now that I draw it out! Thanks!

i can photoshop the colors in if you want. lol
Group 1 turned out decent, if you pretend ... you can see the colors.

I don't think the dot you can see in the photo now was there when we first got the bands. I think when we were measuring we made some marks on there and they are now showing up in the photo. There shouldn't be any color between green and violet, so I don't think there is supposed to be a dot there. At the top, we do have a dot and no color, but we just assume that it was supposed to be blue and use it. The only ones we did not use were the bottom 2 - red and blue since they did not show up and don't have dots.

I also photoshopped mine a little to make it brighter.

It does look like group 5 had another pencil dot. It wasn't supposed to be there - unless somebody was marking where actin appears. That makes sense.