Depression threads

**Shiroikage** · 2012-05-30 14:37

Originally Posted by Chun Liroy

Yes, spin column. I just don't get it either, have no idea what I'm doing wrong.

Buffer PB x5 > 750ul PE > dry spin > 30ul EB

it's been like a year since i did one, but this all sounds right (from what i remember), might have did an extra PB wash
do you have a nanodrop/nanovue or something that you can quickly check whether it's getting lost at any of the steps? i'd look at the eluate @ each step if it's not working

**Chun Liroy** · 2012-05-30 14:38

First 2 times I got bands that were too dim for sequencing, This time nothing. ffff

**Shiroikage** · 2012-05-30 14:41

Originally Posted by Chun Liroy

First 2 times I got bands that were too dim for sequencing, This time nothing. ffff

like i said, look at the eluate at each step
if you're losing it, it's gotta be going somewhere (or just degrading, I guess)

**Chun Liroy** · 2012-05-30 14:41

Originally Posted by Shiroikage

it's been like a year since i did one, but this all sounds right (from what i remember), might have did an extra PB wash
do you have a nanodrop/nanovue or something that you can quickly check whether it's getting lost at any of the steps? i'd look at the eluate @ each step if it's not working

I'm chalking it up to human error right now... which frustrates me more considering how fucking hard is it to pipet things. A monkey could do it brah.

**Mertron** · 2012-05-30 14:42

nerds

**Shiroikage** · 2012-05-30 14:42

make sure you're pipetting the EB onto the center of the column (but not hitting it) & maybe let it sit for a minute or 2

**Shiroikage** · 2012-05-30 14:43

Originally Posted by Spaghetti

nerds

this is like the definition of ultranerd

**Chun Liroy** · 2012-05-30 14:51

Originally Posted by Shiroikage

(but not hitting it)

I think that was the problem, so I took the spin column part and re-ran elution to see if it would work... we'll see

**Charismatic** · 2012-05-30 14:53

Not hitting it? I didn't think that'd be a problem for you.

**Chun Liroy** · 2012-05-30 14:56

Originally Posted by Charismatic

Not hitting it? I didn't think that'd be a problem for you.

Not sure you're in the position to talk.

**Howie Roary** · 2012-05-30 14:56

maybe he's in a position where it's difficult to talk

Spoiler: show

**Chun Liroy** · 2012-05-30 15:24

Welp, didn't work guess today was just a giant waste of time.

**Howie Roary** · 2012-05-30 15:25

um... CPR is pretty easy. I took a course for a hundred bucks, learned it in 8 hours

**Charismatic** · 2012-05-30 15:54

Originally Posted by Chun Liroy

Not sure you're in the position to talk.

And you'll never be sure. Fortunately I will.

**Howie Roary** · 2012-05-30 15:58

will?

you mean am?

**Howie Roary** · 2012-05-30 15:58

VIRGIN FIGHT

**Chun Liroy** · 2012-05-30 16:01

Originally Posted by Howie Roary

VIRGIN FIGHT

Notrly.

**Howie Roary** · 2012-05-30 16:01

hymen or it didn't happen

**Charismatic** · 2012-05-30 16:03

Originally Posted by Charismatic

And you'll never be sure. Fortunately I will.

Originally Posted by Howie Roary

will?

you mean am?

Originally Posted by Howie Roary

VIRGIN FIGHT

Oh, you got dat virginity mah nigga?

**Howie Roary** · 2012-05-30 16:08

no, sadly I am much too slutty

Thread: Depression threads

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