Re: [DIYbio] DNA quantification instruments

GUIs in python are pretty easy too. Here's some old code I wrote to plot spectrometer data in python:
https://github.com/nmz787/open-spectrometer/blob/master/desktop-software/openSpectrometer.py

I've been working with wxpython lately for GUIs, and would have to look at the pygtk forums and bug tracker to see if it would be worth switching to wxpython or not in any future updates/forks of this. I kind of forgot about it when I was looking at GUI toolkits a few months ago.

On Jul 31, 2014 7:14 PM, "Nathan McCorkle" <nmz787@gmail.com> wrote:

Excel read/write in python is easy.

On Jul 31, 2014 6:41 PM, "Cory Tobin" <cory.tobin@gmail.com> wrote:
On Thu, Jul 31, 2014 at 5:10 PM, John Griessen <john@industromatic.com> wrote:
> What format of data do your colleagues want?  Or, (even worse), is there a
> specific
> vendor and app they use?

The Nanodrop software can export your data as an Excel spreadsheet.  I
know plenty of people who would whine for days if this was no longer
an option.  Not because Excel is particularly useful but because
that's the way they've done it for years.  Some biologists are
obstinate.

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Re: [DIYbio] DNA quantification instruments

Excel read/write in python is easy.

On Jul 31, 2014 6:41 PM, "Cory Tobin" <cory.tobin@gmail.com> wrote:
On Thu, Jul 31, 2014 at 5:10 PM, John Griessen <john@industromatic.com> wrote:
> What format of data do your colleagues want?  Or, (even worse), is there a
> specific
> vendor and app they use?

The Nanodrop software can export your data as an Excel spreadsheet.  I
know plenty of people who would whine for days if this was no longer
an option.  Not because Excel is particularly useful but because
that's the way they've done it for years.  Some biologists are
obstinate.

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Re: [DIYbio] DNA quantification instruments

On Thu, Jul 31, 2014 at 5:10 PM, John Griessen <john@industromatic.com> wrote:
> What format of data do your colleagues want? Or, (even worse), is there a
> specific
> vendor and app they use?

The Nanodrop software can export your data as an Excel spreadsheet. I
know plenty of people who would whine for days if this was no longer
an option. Not because Excel is particularly useful but because
that's the way they've done it for years. Some biologists are
obstinate.

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Re: [DIYbio] Paper Acces

Hello, I need pdf Collaborative study of methods for solvent retention capacity profiles (AACC methods 56-11 of Gaines 2000.

Thanks you

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Re: [DIYbio] DNA quantification instruments

On 07/31/2014 05:52 PM, Cory Tobin wrote:
> It doesn't report data to a computer so you have to resort to
> recording your data into your notebook with a pen. I don't mind but I
> know this would be a deal breaker for some of my colleagues.

What format of data do your colleagues want? Or, (even worse), is there a specific
vendor and app they use?

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Re: [DIYbio] DIY Protein Separation

wouldn't you use protein SDS gel electrophoresis to separate the
protein? or is that too analytical and you want preparative (i.e. you
need a lot of your pure protein)?

I guess how much protein do you need? I seem to recall Josiah posting
on biocurious that he was cleaning up a F/H-PLC to try and get
working... so I presumed he has some experience with them. I've some
HPLC and GC once in a class, and according to wikipedia ion exchange
chromatography is also considered FPLC sometimes if used for protein,
so when I used the Carolina kit for GFP I guess I was doing FPLC :)

These terms are all a bit overlapping, kinda like the act of walking,
whether you have sandals or boots on. A lot of similarity, but
differences around the edges.

normal phase vs reverse phase (is the buffer (mobile phase) polar or
the resin (solid phase) polar?). You can also have different phases
than solid and liquid. You could say that your lungs filter oxygen and
repel CO2, that would probably be something like gas to liquid to
solid... but that might be stretching it for an /easy/ example :)


On Thu, Jul 31, 2014 at 6:09 AM, Mega [Andreas Stuermer]
<masterstorm123@gmail.com> wrote:
> Hi!
> Again to my question, there seem to be little guys dealing with FPLC here.
>
> https://groups.google.com/forum/#!topic/diybio/24PZTzije98
>
> Is there a way to do this cheaply at my university, without buying an FPLC
> machine?
>
> Thanks god those berberine derivatives are all fluorescent. So detecting
> will not be a problem.
>
> What we want to do is: preparing a cell protein extract. Separate those
> proteins. Add the fractions to berberine, let it incubate, and do
> photometry see which turns it into the derivative we want.
>
> Best,
> Andreas
>
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Re: [DIYbio] DNA quantification instruments

I recently bought an old Eppendorf Biophotometer. I have access to a
Nanodrop at my university's campus and the Biophotometer reports DNA
concentrations within one 1 or 2 ng/uL of what the Nanodrop reports.
There are a few downsides to the Biophotometer, though.

1) It only reads at fixed wavelengths (230,260,280 and a few others)
so you can't get a full spectrum measurement like on the Nanodrop.

2) By itself it does not handle microliter volumes. It takes a
cuvette like usual. But it can also handle the Hellma TrayCell or the
Eppendorf uCuvette G1.0. Those are sold separately but allow for
using 1-2uL samples. These devices fit into the cuvette receptacle
and focus the light through a small point where the sample sits. I
have a Hellma TrayCell and it works just fine. It's a little more of
a pain to clean out the old sample compared to the Nanodrop but I
don't mind.

3) It doesn't report data to a computer so you have to resort to
recording your data into your notebook with a pen. I don't mind but I
know this would be a deal breaker for some of my colleagues.

4) The TrayCells are hard to find (occasionally one will show up on
ebay) and the uCuvette G1.0 costs around ~2,000 USD.

The positives are

1) Way cheaper than a Nanodrop.

2) Works just as well as a Nanodrop.

3) Can also do OD measurements so you don't need a separate device.

-cory

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Re: [DIYbio] Re: diy electroporation so far?

On 7/31/14 8:21 AM, Nathan McCorkle wrote:


On Jul 30, 2014 10:45 PM, "Jonathan Cline" <jncline@gmail.com> wrote:

> I guess a piezo starter might work but I would think it would be rather imprecise.  Nature is imprecise too.  But you want to ensure a good result and/or a good yield.

Months ago I got no/little response when I posted a paper theorizing lightning strikes as potential for horizontal gene transfer (i.e. electroporation)

Was that an earth shattering idea?   (No pun intended.)

## Jonathan Cline  ## jcline@ieee.org  ## Mobile: +1-805-617-0223  ########################    
 

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SAS Data Conversion Specialist with Mainframe Experience (Financial experience) - TX

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Urgent requirement : Job Opportunity- Hadoop Admin In Hadoop Admin

 

Hello,                       

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Re: [DIYbio] Re: Max resistor voltage

On 07/31/2014 12:13 PM, Cathal (Phone) wrote:
> I like the graphite/paper concept!
+1

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Re: [DIYbio] Re: Max resistor voltage

On 07/31/2014 11:26 AM, Simon Quellen Field wrote:
> the idea I like best is to draw two lines on paper with a pencil and let a drop of critters soak into the paper. You can make the
> lines quite close together, and graphite isn't toxic. Then cut the paper and drop it into some nutrient broth.
>
> No cleaning electrodes.
> You can control the volume quite easily.
> Nothing dangerous.

Hmm... that idea even upgrades from DIY to a low cost product! How about ink jet printed conductors
with some large lands that spring contacts connect to on your electroporator-with-safety-features?
The paper could come from the clean and pure sources used for old school analytic filter paper.

It could come in sealed sterile packs. Hey Nathan -- want to do some testing of that starting with
hand drawn graphite papers? The extra volume of critters outside the lines may not matter.
For apps where a tiny volume is all you have the paper could be trimmed away just outside the conductive lines
and dip it in a microdrop of bugs to load your bugs into the "zap chamber"

Fantastic lateral thinking Simon!

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Re: [DIYbio] Re: Max resistor voltage

I expect you'd get greater efficiency with steel as copper ions are pretty cytotoxic. I like the graphite/paper concept!

On 31 July 2014 17:26:37 GMT+01:00, Simon Quellen Field <sfield@scitoys.com> wrote:
You can get copper tape at a hardware store. It is used to keep snails out of gardens, and to stick around windows for burglar alarms.

But the idea I like best is to draw two lines on paper with a pencil and let a drop of critters soak into the paper. You can make the lines quite close together, and graphite isn't toxic. Then cut the paper and drop it into some nutrient broth.

No cleaning electrodes.
You can control the volume quite easily.
Nothing dangerous.


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On Thu, Jul 31, 2014 at 8:26 AM, Nathan McCorkle <nmz787@gmail.com> wrote:

My last trials with a piezoelectric sparker used two pieces of adhesive-backed aluminum foil on a glass microscope slide, with some wax pencil for a  hydrophobic 'corral' between the two foil strips. Seemed like it should work as long as aluminum ions weren't toxic to the cells.

I really think the next step is to add a diode (LED), and a small cap/inductor would be the next step.

On Jul 31, 2014 7:29 AM, "John Griessen" <john@industromatic.com> wrote:
On 07/30/2014 05:15 PM, Simon Quellen Field wrote:
You might want to simply place two wires very close to one another inside the cuvette (or Petri dish -- we don't need the cuvette
anymore) and hit the igniter a couple dozen times.

This idea could yield some safer lower powers to electroporate with.  The concept is to make the
field strength quantified by the volume between the electrodes and not worry with it being a cuvette.
It could be just small zones of metal separated by a distance.

Then you zap a droplet in any container.

Drawbacks include:
1. needing to reuse such electrodes and so having to clean them.
2. The amount or number of cells affected will vary as the volume you put the electrodes in varies.
3. electrodes are sticking out where they could be dangerous.

So, with the drawbacks considered, I still like the idea of making teeny cuvettes to get the needed currents lower.
They would be used inside an insulated interlocked covered box.

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Sr. ETL/Teradata Developer Available Immediately

Hello,

 

Hope you are doing well.

 

We have ETL/Teradata Developer candidate available in CA. She is available immediately & looking for projects in CA.

Kindly let me know if you have any DIRECT CLIENT/Tier 1 Vendor’s opening where we can submit her.

 

 

SUMMARY:

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·          Experience in all phases of Data warehouse development from Requirements, analysis, design, development, testing and post production support.

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·         Independent, Self-starter,Hadoop, enthusiastic team player with strong adaptability to new technologies.

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ETL Tools

Informatica 6.x/7.x/8.x/9.1, Data Stage

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GUI

.Net Custom development, Business Objects, Micro Strategy

Operating Systems

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Domain Knowledge

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Re: [DIYbio] Re: Max resistor voltage

You can get copper tape at a hardware store. It is used to keep snails out of gardens, and to stick around windows for burglar alarms.

But the idea I like best is to draw two lines on paper with a pencil and let a drop of critters soak into the paper. You can make the lines quite close together, and graphite isn't toxic. Then cut the paper and drop it into some nutrient broth.

No cleaning electrodes.
You can control the volume quite easily.
Nothing dangerous.


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On Thu, Jul 31, 2014 at 8:26 AM, Nathan McCorkle <nmz787@gmail.com> wrote:

My last trials with a piezoelectric sparker used two pieces of adhesive-backed aluminum foil on a glass microscope slide, with some wax pencil for a  hydrophobic 'corral' between the two foil strips. Seemed like it should work as long as aluminum ions weren't toxic to the cells.

I really think the next step is to add a diode (LED), and a small cap/inductor would be the next step.

On Jul 31, 2014 7:29 AM, "John Griessen" <john@industromatic.com> wrote:
On 07/30/2014 05:15 PM, Simon Quellen Field wrote:
You might want to simply place two wires very close to one another inside the cuvette (or Petri dish -- we don't need the cuvette
anymore) and hit the igniter a couple dozen times.

This idea could yield some safer lower powers to electroporate with.  The concept is to make the
field strength quantified by the volume between the electrodes and not worry with it being a cuvette.
It could be just small zones of metal separated by a distance.

Then you zap a droplet in any container.

Drawbacks include:
1. needing to reuse such electrodes and so having to clean them.
2. The amount or number of cells affected will vary as the volume you put the electrodes in varies.
3. electrodes are sticking out where they could be dangerous.

So, with the drawbacks considered, I still like the idea of making teeny cuvettes to get the needed currents lower.
They would be used inside an insulated interlocked covered box.

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RE: [DIYbio] DNA quantification instruments

I own a qubit and its indispensible. The strand specific DNA stain coupled with the tiny footprint makes it a fantastic deal. Only catch is that it won't tell you things like salt conc. so you can't tell how pure your sample is.

Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC
Plant Biotech R&D

From: Nathan McCorkle
Sent: ‎7/‎31/‎2014 11:30 AM
To: diybio
Subject: Re: [DIYbio] DNA quantification instruments

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Re: [DIYbio] DNA quantification instruments

The nanodrop /is/ just a plain old spectrometer, with two fiber optic cables leading to and from the microdrop stage. Search Google for TakeItApart nanodrop to see pics of the inside.

On Jul 31, 2014 3:00 AM, "jarlemag" <jarle.pahr@gmail.com> wrote:
Hello. As mentioned in a few previous posts I'm shopping for lab equipment, and one of the pieces I'm missing is some kind of equipment for quantifying DNA. I have considered several alternatives, and would like to ask about your experiences. To specify my needs a bit more, I am primarily interested in doing DNA concentration measurements of samples (PCR products) to be sent for sequencing. So preferably, the method should not require a too large sample volume.

Previously, I have used solely the NanoDrop (ND-1000) for measuring DNA concentrations. It's a nifty instrument and I'd love to have one but the prices I have seen have been in the $10 000 range, which is far above my current budget. Because it is a relatively new product, there are also few used units being sold. The main attraction of the NanoDrop is the microliter sample volume and ease of use. The main criticism I have seen of the ND, and which I agree with, is poor accuracy at low DNA concentrations with too high DNA concentrations being reported. However, the absorbtion curve being presented visually for each measurement at least makes it possible to evaluate how much trust to put in each measurement. There are several NanoDrop "clones" available too, such as Quawell (http://www.quawell.com/), but these are still priced at $5000 or so, and I'm unsure of the quality.

Although I'm unlikely to purchase one in the near future, I'd still like to hear your opinions on NanoDrop and other microliter-spectrometers. Are they worth the expense?

An instrument which I have seen touted as a cheaper and more accurate alternative to the NanoDrop is the Qubit Fluorometer.(http://www.lifetechnologies.com/no/en/home/brands/product-brand/qubit.html?cid=fl-qubit). Especially at low DNA concentrations, the Qubit seems to be more useful. The downsides are larger sample volume (up to about 20 uL needed), some sample preparation needed, and the need to buy instrument-specific consumables. Still, this instrument can be had for about 2-3000 $USD, which is affordable and would seem to give good value if the Qubit is as accurate as the advertisments suggests.

Have any of you used the Qubit or similar fluorescent-based DNA quantification instruments? What has your experience been?

The third alternative I am aware of is a plain old spectrometer. I also want something for measuring optical density of bacterial cultures, so a spectrometer is something I'd like or need for that purpose anyway. My main concern with using one for DNA concentration measurements is the need for (much) larger sample volumes than with NanoDrop or Qubit. Rather than buying a large, heavy and expensive all-round spectrometer, I am considering buying an Eppendorf Biophotometer (http://www.eppendorf.com/int/index.php?pb=63cfedee26795e18&action=products&contentid=1&catalognode=53772) (That's the newest model, but there are used older models for sale much cheaper). This seems to give a good all-round package for OD600, DNA/RNA and protein concentrations at smaller size and lower price. There are also small, cheap generalized spectrometers of Chinese make available, but I am very unsure of the quality of these...

Do you any of you have experience with the Eppendorf Biophotometer/Biospectrometer series or other spectrometers for measuring DNA concentrations? How much work goes into making standard curves and calibration? What is the minimum practical sample amount?


And finally, for those who have used several of the methods above: What do you recommend as the "go to" method for measuring DNA concentrations for a new DIYBio laboratory?

Looking forward to hear your opinions!

Best regards,
Jarle Pahr

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