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hogthehedge

Member Since 11 Apr 2012
Offline Last Active Dec 03 2012 12:21 PM

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ascacioc
06 Sep 2012 - 18:48
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Topics I've Started

Chromosome mapping problem

02 November 2012 - 08:15 AM

Hi everyone,

I'm trying to work this problem on contigs in Lehninger's Biochemistry and am having no luck. I've tried arranging the fragments per size, and counting overlaps, but I feel like I'm missing some crucial bit of logic that will switch on the bulb!

"A group of overlapping clones, designated A through F, is isolated from one region of a chromosome. Each of the clones is seperately cleaved by a restriction enzyme and the pieces resolved by agarose gel electrophoresis, with the results shown in the figure below. There are nine different restriction fragments in this chromosomal region, with a subset appearing in each clone. Using this information, deduce the order of the restriction fragments in the chromosome"

I've attached the gel image that the problem provides.

Any ideas, tips, or even the whole solution ;-) will be much appreciated~!

Thanks,

Hogthehedge

Image-sharing etiquette

05 September 2012 - 12:22 PM

Hi everyone,

I need some advice on what to do. I've been working in this lab for a while, and I've made some beautiful images and time-lapse captures. My time here is done, I'm moving on to another lab. My PI wants these images before I leave. How can I tactfully ask for credit for them, without coming across as too pushy? Not academic credit. I mean, if he puts up those time-lapse captures on the lab website, I'd really like it to say I made them. I know I made them using his microscope and his facilities, but I still made them. I'm sort of tempted to just inscribe my name at the bottom of the videos. Is that OK etiquette-wise? Or does he automatically own all my work?

Ampicillin resistance selector

01 August 2012 - 03:43 AM

Hi everyone,

Do you know which antibiotic to use to select mammalian cells expressing a plasmid with Ampicillin resistance?
I use Geneticin for Kanamycin-resistance conferring plasmids. Would the same work?

Regards,
HogtheHedge

Time on ice before prepping

14 July 2012 - 01:46 AM

Hi everyone,

I was wondering if it was OK to put my culture in the 4C fridge for an entire day before I prepped it. Would my maxiprep still work out OK? Any idea how it will affect the plasmid quality/yield?

Regards,
hogthehedge

MW of PEG for Purification of DNA

28 June 2012 - 03:18 AM

Hi,

I'm following the Cold Spring Harbor labs' protocol for PEG purification of DNA. The PEG-MgCl2 solution they describe is as follows (from http://cshprotocols....06/1/pdb.rec382)

40% (w/v) polyethylene glycol (PEG 8000)
30 mM MgCl₂

Dissolve 40 g of PEG 8000 in a final volume of 100 ml of 30 mM MgCl₂. Sterilize the solution by passing it through a 0.22-μm filter, and store it at room temperature.

However, I only have PEG of MW 3350 in the lab, not PEG of MW 8000. Will it still work with PEG 3350?
Why/why not? I don't know how PEG works to precipitate DNA and I would really appreciate it if someone who knew explained it to me.

Thanks,

HogtheHedge

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