Re: [DIYbio] Paralens fluorescence microscope objective for Hoechst and DAPI?

Abizar,

Thank you - yes I'd be very interested in trying the setup you suggest - please can you post an image? I have Zeiss Standard 25 binocular microscope. The key thing is that I need a sharp fluorescence image with a 100x oil objective.

- Is the setup you suggest compatible with this type of objective?
- Where can I get the required parts and how do I make them fit my microscope (LED, filters, camera)?
- Can I acquire LED lighting that will provide for UV illumination (for use with DAPI, Hoechst)?

Roninlaw,

The price of the core Paralens Advance system with a single objective lens type is approximately £900 including VAT.

Thanks guys,

Mike

On Friday, June 23, 2017 at 11:15:21 PM UTC+1, Roninlaw wrote:
Price?

On Fri, Jun 23, 2017 at 2:25 PM, Mike <alanmb...@gmail.com> wrote:
Hello all,

As a cheap alternative to a fluorescence microscope for my home lab, I am considering the Paralens Advance screw-on objective lens.

(Please see page 9 of the PDF for an illustration of light spectra)

The Paralens produces excitation at 385-480nm and emission at 480nm (as far as I can tell, please see PDF above!). [b]Question[/b]: Will this device work with Hoechst 33258 and DAPI stain (spectra as below)?

- Hoechst 33258: excitation: 352, emission: 461
- DAPI: excitation: 358, emission: 461

Thank you,

Mike

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Re: [DIYbio] Paralens fluorescence microscope objective for Hoechst and DAPI?

An alternative is to place your illuminating led and filter on one eyepiece of a binocular head. And a filter and camera on the other eyepiece. The led beam is semi focused on the specimen through the objective. Fluorescence light goes up the optical path and is divided equally into the eyepiece though only the camera sees the fluorescence image.
Can send you an image of this setup if it helps.

On Jun 23, 2017 2:25 PM, "Mike" <alanmbeattie@gmail.com> wrote:
Hello all,

As a cheap alternative to a fluorescence microscope for my home lab, I am considering the Paralens Advance screw-on objective lens.

(Please see page 9 of the PDF for an illustration of light spectra)

The Paralens produces excitation at 385-480nm and emission at 480nm (as far as I can tell, please see PDF above!). [b]Question[/b]: Will this device work with Hoechst 33258 and DAPI stain (spectra as below)?

- Hoechst 33258: excitation: 352, emission: 461
- DAPI: excitation: 358, emission: 461

Thank you,

Mike

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Re: [DIYbio] Paralens fluorescence microscope objective for Hoechst and DAPI?

Price?

On Fri, Jun 23, 2017 at 2:25 PM, Mike <alanmbeattie@gmail.com> wrote:
Hello all,

As a cheap alternative to a fluorescence microscope for my home lab, I am considering the Paralens Advance screw-on objective lens.

(Please see page 9 of the PDF for an illustration of light spectra)

The Paralens produces excitation at 385-480nm and emission at 480nm (as far as I can tell, please see PDF above!). [b]Question[/b]: Will this device work with Hoechst 33258 and DAPI stain (spectra as below)?

- Hoechst 33258: excitation: 352, emission: 461
- DAPI: excitation: 358, emission: 461

Thank you,

Mike

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[DIYbio] Paralens fluorescence microscope objective for DAPI and Hoechst?

Hello all,

As a cheap alternative to a fluorescence microscope for my home lab, I am considering the Paralens Advance screw-on objective lens.

http://www.druckerdiagnostics.com/media/wysiwyg/paralens-advance/PLA_Manual_No_Spreads_.pdf
(Please see page 9 of the PDF for an illustration of light spectra)

The Paralens produces excitation at 385-480nm and emission at 480nm (as far as I can tell, please see PDF above!). [b]Question[/b]: Will this device work with Hoechst 33258 and DAPI stain (spectra as below)?

- Hoechst 33258: excitation: 352, emission: 461
- DAPI: excitation: 358, emission: 461

Thank you,

Mike

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[DIYbio] Paralens fluorescence microscope objective for Hoechst and DAPI?

Hello all,

As a cheap alternative to a fluorescence microscope for my home lab, I am considering the Paralens Advance screw-on objective lens.

http://www.druckerdiagnostics.com/media/wysiwyg/paralens-advance/PLA_Manual_No_Spreads_.pdf
(Please see page 9 of the PDF for an illustration of light spectra)

The Paralens produces excitation at 385-480nm and emission at 480nm (as far as I can tell, please see PDF above!). [b]Question[/b]: Will this device work with Hoechst 33258 and DAPI stain (spectra as below)?

- Hoechst 33258: excitation: 352, emission: 461
- DAPI: excitation: 358, emission: 461

Thank you,

Mike

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Looking for Sr. Python Developer , WA

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Re: [DIYbio] Re: SLICE -- Seamless Ligation Cloning Extract... or, using lambda phage homologous recombination in-vitro

Probably because it requires an extra PCR reaction. 

The separate SLiCE "enzyme mix" you refer to costs nothing, and with about 2 hrs of prep work one can make enough for 10,000+ reactions. PCRs are not free, often require troubleshooting and can be time consuming. The fewer PCRs one has to do the better in my book. 

Granted not all cloning methods work equally well for all situations, but for standard insertion of an insert into a linearized vector it's pretty hard to beat the convenience, cost, efficiency and accuracy of recombinase-expressed cell extract SLICE methods. I suspect those making $$ on Gibson reagents are working hard to ensure these methods don't become commercialized. 



On Wednesday, May 3, 2017 at 11:10:57 PM UTC-7, Michael Crone wrote:
Why not just use CPEC instead of requiring a separate enzyme mix just for cloning? Phusion works absolute wonders and I've never had any problem creating large plasmids.

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Re: [DIYbio] Re: Gibson vs SLiCE (single part recommendations)

Thanks for your input Cihan. I would also be particularly curious to see how the plasmid-based recombinase extract works compared to PPY/genome-based.

The Motohashi papers on using traditional extracts are quite interesting. Thanks for sharing. I've read through his original paper (https://bmcbiotechnol.biomedcentral.com/articles/10.1186/s12896-015-0162-8) and he certainly gets very different numbers for non-recombinase extract than Zhang did, in both cloning efficiency (up to 2-orders of magnitude better) and ideal homology overlap length (20bp compared to 50bp from Zhang). Reasons for the discrepancy are not well discussed. The reported cloning accuracy when combining two inserts into a vector was kind of low as well. In our lab now with the PPY strain we are putting in 1-4 fragments at a time with nearly 100% accuracy, and with good enough efficiency to assemble large libraries. 

This is not to say the methods of using traditional cell extracts don't work well or aren't perfectly suitable for many labs and many applications...I suppose my point is that I am a believer at this point that making extracts from cells expressing recombinase (whether PPY or plasmid based) really does make a difference. And when students clone more efficiently, projects move forward faster. 

One of these days I'll get around to doing a more systematic side-by-side comparison of using extracts with and without recombinase.

Just my two cents. ;)




On Tuesday, June 20, 2017 at 1:13:07 AM UTC-7, Cihan AYDIN wrote:
I also got an MTA for the PPY strain but the shipping costs (for cold chain) was too much for me so I opted for getting a pKD46-deriven plasmid (pREDIA from Addgene). The only difference is in PPY gam and redb is integrated into the DH10B genome and reda in plasmid whereas in pKD 46 it is on the plasmid. I really would like to make a side-by-side comparison for the PPY and pKD46/DH10B in my lab.

Also there is this japanese guy Motohashi who does SLiCe from regular bacteria extracts (recA-) and has data on its efficiency with 15-19 bp overlaps between parts (https://link.springer.com/protocol/10.1007%2F978-1-4939-6472-7_23). He is using JM109 as his primary strain and uses a few more alternatives as far as i remember.

On the note, Gibson is too pricey for any lab who's on a budget, especially DIY labs and labs in developing countries. SLiCe and even CPEC has become better alternatives for synthetic biology and even simple cloning reactions - basically your only cost is bacterial reagents and primers (and PCR reagents in some cases).

On a last note, also read this article - www.ncbi.nlm.nih.gov/pubmed/26463009 - where they combine CRISPR with Lambda phage recombination system to efficiently modify e. coli genome.

Cheers!

C

On Sunday, June 18, 2017 at 6:07:48 AM UTC+3, Rob C wrote:
Not too long ago we obtained the PPY strain as a kind gift from the original makers (Zhang at Albert Einstein). 

In our hands making extracts from this strain works WAY better than traditional DH10b, DH5alpha, etc (using the CelLytic lysis B reagent...it's cheap for what you get). 

Although I don't have exact numbers to report, we have done several tests side by side with Gibson, and PPY based SLICE gives at least equal transformation efficiency. Accuracy is equally as good, and in some isolated cases it has been better. 

After several month of heavy cloning, our lab has now dropped purchase of all Gibson reagents. 

I'm convinced that if more people implement the PPY-based SLICE cloning in their lab, Gibson sales will go out of business. 



On Thursday, March 16, 2017 at 12:32:35 PM UTC-7, Rob C wrote:
Bryan, thank you very much for sharing your experiences. Sounds like you did all the right controls to get a proper assessment of the system. Logically it is a little surprising that there's no carry over of plasmid, but of course very good news and just goes to show the best way to answer a question is to just do the experiment. I tend to see too many people over thinking things to a point where they will convince themselves something is not feasible before they've even tried it, so the project stops before it even begins. :)

Cheers.

On Monday, March 13, 2017 at 11:57:49 AM UTC-7, Bryan Jones wrote:
I would imagine there is some risk of that, but I haven't seen that happen in my experiments. I carried out a negative control using just SLiCE extract with competent cells (No insert DNA), and didn't get any colonies on amp plates. Also the colonies I sequenced from my tests were all correct, no pKD46 carryover. It might be that DNAse breaks all the DNA from the SLiCE cells. Normally when you do a plasmid prep, you add EDTA to prevent this, but no EDTA was added here. I'm not positive why, but empirically, I can say that it does not seem to be an issue. 

On Tue, Mar 7, 2017 at 6:16 PM RBC <rco...@gmail.com> wrote:
Is there any worry that the amp resistant pKD46 plasmid will be extracted into the SLiCE lysate, in which case this plasmid will carry through to the final transformation? To me it would seem you can't use it to make amp resistant vectors..or maybe I'm missing something. Thanks for the clarification.

RC


On Friday, October 21, 2016 at 11:22:11 AM UTC-7, Bryan Jones wrote:
In my most recent test, without pKD46, I got 10 colonies from the SLiCE reaction, which was the same as the no SLiCE control. SLiCE with pKD46 resulted in 50-60 colonies. For comparison, NEBuilder resulted in about 100.



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[DIYbio] Re: Human DNA sequencing

Thanks,
This will be helpful for comparison.

On Saturday, June 17, 2017 at 8:08:53 AM UTC-5, Christopher Monaco wrote:
You can check out the 1000 genomes project, http://www.internationalgenome.org/ you can download genomic data from already assembled and annotated genomes all the way down to the raw sequencing data. I'm not sure if the metadata contains info on disease states of the inviduals, but you can always tie mutations back to known diseases.

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Re: [DIYbio] Gel Red questions

For those interested, I came across this dye, based on a food additive.
http://www.tintorang.com/

This alone caught my attention:

tinto rang claims to be 100% safest, as it is a plant based food grade product, with zero toxicity, mutagenicity and is non-hazardous.

It will solve the issues with EtBr. 

http://www.tintorang.com/Nucleic-Acid-Gel-Stain/tinto-rang-safe-nucleic-acid-gel-stain/safe-food-grade-nucleic-acid-DNA-RNA-Fluorescent-Dyes.html

-Ravi


On Sunday, April 30, 2017 at 11:39:37 PM UTC+5:30, John Ladasky wrote:
SYBR Safe and Thiazole Orange are very similar, but not identical.  So is Thiazole Orange an acceptable substitute for the more expensive SYBR Safe in gel staining?  If so, why isn't everyone using it?

Thiazole Orange is membrane-permeable.  It was first developed to label RNA in intact reticulocytes.  I'm pretty sure that Thiazole Orange also binds DNA.  All DNA intercalating dyes are suspected carcinogens, for obvious reasons.

One way that SYBR Safe might be "safer" than Thiazole Orange is if the former was membrane-impermeant.  Is it?  I doubt it.  If anything, I would expect SYBR Safe to be slightly more membrane-permeant, because SYBR Safe has a propyl group where Thiazole Orange has a methyl group.  However, that structural change might also reduce toxicity.

I just had a look at an MSDS for each compound, and I didn't see any obvious differences.  



On Thursday, April 27, 2017 at 1:49:25 PM UTC-7, Dennis Oleksyuk wrote:
It is much cheaper here if you buy 5G at a time.

On Wed, Apr 26, 2017 at 5:06 PM Mega [Andreas Stuermer] <masters...@gmail.com> wrote:







On Sunday, April 23, 2017 at 10:03:09 PM UTC+2, John Ladasky wrote:


On Tuesday, March 12, 2013 at 12:03:42 PM UTC-7, Nathan McCorkle wrote:
Yeah, that's why big companies and universities don't switch.

EtBr is molecularly known (no one really knows exactly what the gelRed
and sybrSafe molecules are, we know what classes they're in or one
that was described in one patent, etc)

I'm pretty sure that this information cannot be kept secret.  To obtain a chemistry patent, and to publish an accurate MSDS, the structure of the novel active compound must be known and disclosed.

I remember that SybrSafe is in the cyanine dye family, a descendant of Thiazole Orange.  If you look at the Wikipedia article for SybrSafe, a structure is published, and it sure looks like a cyanine dye to me.

 
Wikipedia entries are also available for GelGreen and GelRed.  They look like acridine orange and ethidium dimers, respectively, with a peptide-ether linker between the individual fluorophores.


The idea of making ethidium dimers is not new.  An ethidium dimer was available in 1990.  It's still available.  The linker differs from the GelRed linker, but otherwise the two molecules are quite similar.



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[DIYbio] In vivo T4 DNA ligase expression.

Hi all, 

I often do simple cloning of unmodified PCR products using a suicide vector. I wonder if anyone has tried to express the T4 DNA ligase in vivo so that the 
ligation could happen in vivo? 

I have only seen this paper by Ren et al https://www.ncbi.nlm.nih.gov/pubmed/9305776

They express T4 ligase from a plasmid and they manage to recircularize linear plasmids efficiently
in a way that is dependent on T4 ligase expression. 

They do some experiments with double digested vectors where they cut out an internal part of a marker.
They get this marker back in 9 out of 50 cases (if I understood correctly, Table 4 ) for a recABC background.

Since there are two orientations, this means that 18 out of 50 clones are recombinant. 
With some kind of counter selection against empty clones (I use the pCAPs vector, works very well) and loading more insert, the frequency should be 
so high that you do not need to look at perhaps 3-5 clones.

Did anyone try this or something similar? This would be great for DIY, since there would be no need to purify the ligase. 

/bjorn



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Re: [DIYbio] Re: Gibson vs SLiCE (single part recommendations)

I also got an MTA for the PPY strain but the shipping costs (for cold chain) was too much for me so I opted for getting a pKD46-deriven plasmid (pREDIA from Addgene). The only difference is in PPY gam and redb is integrated into the DH10B genome and reda in plasmid whereas in pKD 46 it is on the plasmid. I really would like to make a side-by-side comparison for the PPY and pKD46/DH10B in my lab.

Also there is this japanese guy Motohashi who does SLiCe from regular bacteria extracts (recA-) and has data on its efficiency with 15-19 bp overlaps between parts (https://link.springer.com/protocol/10.1007%2F978-1-4939-6472-7_23). He is using JM109 as his primary strain and uses a few more alternatives as far as i remember.

On the note, Gibson is too pricey for any lab who's on a budget, especially DIY labs and labs in developing countries. SLiCe and even CPEC has become better alternatives for synthetic biology and even simple cloning reactions - basically your only cost is bacterial reagents and primers (and PCR reagents in some cases).

On a last note, also read this article - www.ncbi.nlm.nih.gov/pubmed/26463009 - where they combine CRISPR with Lambda phage recombination system to efficiently modify e. coli genome.

Cheers!

C

On Sunday, June 18, 2017 at 6:07:48 AM UTC+3, Rob C wrote:
Not too long ago we obtained the PPY strain as a kind gift from the original makers (Zhang at Albert Einstein). 

In our hands making extracts from this strain works WAY better than traditional DH10b, DH5alpha, etc (using the CelLytic lysis B reagent...it's cheap for what you get). 

Although I don't have exact numbers to report, we have done several tests side by side with Gibson, and PPY based SLICE gives at least equal transformation efficiency. Accuracy is equally as good, and in some isolated cases it has been better. 

After several month of heavy cloning, our lab has now dropped purchase of all Gibson reagents. 

I'm convinced that if more people implement the PPY-based SLICE cloning in their lab, Gibson sales will go out of business. 



On Thursday, March 16, 2017 at 12:32:35 PM UTC-7, Rob C wrote:
Bryan, thank you very much for sharing your experiences. Sounds like you did all the right controls to get a proper assessment of the system. Logically it is a little surprising that there's no carry over of plasmid, but of course very good news and just goes to show the best way to answer a question is to just do the experiment. I tend to see too many people over thinking things to a point where they will convince themselves something is not feasible before they've even tried it, so the project stops before it even begins. :)

Cheers.

On Monday, March 13, 2017 at 11:57:49 AM UTC-7, Bryan Jones wrote:
I would imagine there is some risk of that, but I haven't seen that happen in my experiments. I carried out a negative control using just SLiCE extract with competent cells (No insert DNA), and didn't get any colonies on amp plates. Also the colonies I sequenced from my tests were all correct, no pKD46 carryover. It might be that DNAse breaks all the DNA from the SLiCE cells. Normally when you do a plasmid prep, you add EDTA to prevent this, but no EDTA was added here. I'm not positive why, but empirically, I can say that it does not seem to be an issue. 

On Tue, Mar 7, 2017 at 6:16 PM RBC <rco...@gmail.com> wrote:
Is there any worry that the amp resistant pKD46 plasmid will be extracted into the SLiCE lysate, in which case this plasmid will carry through to the final transformation? To me it would seem you can't use it to make amp resistant vectors..or maybe I'm missing something. Thanks for the clarification.

RC


On Friday, October 21, 2016 at 11:22:11 AM UTC-7, Bryan Jones wrote:
In my most recent test, without pKD46, I got 10 colonies from the SLiCE reaction, which was the same as the no SLiCE control. SLiCE with pKD46 resulted in 50-60 colonies. For comparison, NEBuilder resulted in about 100.



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[DIYbio] Zageno purchasing platform

Hi everyone,

I wanted to share a convenient purchasing platform that we have been using for several months at Boslab.

It's a startup called Zageno. They're constantly increasing their catalog, and if you want something that is not in there you just need to ask them to add it. 

Briefly, advantages are the following:
- They provide Free shipping for all products!! I think this is temporary, but it's been here for a while. We saved a lot of money because we are usually not purchasing products in bulk. 
- You can compare products on their website (cost per reaction, etc.)
- They have a good customer service, you can just throw a list of reagents to them, and they make the quote for you 
- Prices are directly displayed on the website

I hope that helps other DIYbio labs out there.

Best,
Raphael

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Re: [DIYbio] Re: Gibson vs SLiCE (single part recommendations)

Not too long ago we obtained the PPY strain as a kind gift from the original makers (Zhang at Albert Einstein). 

In our hands making extracts from this strain works WAY better than traditional DH10b, DH5alpha, etc (using the CelLytic lysis B reagent...it's cheap for what you get). 

Although I don't have exact numbers to report, we have done several tests side by side with Gibson, and PPY based SLICE gives at least equal transformation efficiency. Accuracy is equally as good, and in some isolated cases it has been better. 

After several month of heavy cloning, our lab has now dropped purchase of all Gibson reagents. 

I'm convinced that if more people implement the PPY-based SLICE cloning in their lab, Gibson sales will go out of business. 



On Thursday, March 16, 2017 at 12:32:35 PM UTC-7, Rob C wrote:
Bryan, thank you very much for sharing your experiences. Sounds like you did all the right controls to get a proper assessment of the system. Logically it is a little surprising that there's no carry over of plasmid, but of course very good news and just goes to show the best way to answer a question is to just do the experiment. I tend to see too many people over thinking things to a point where they will convince themselves something is not feasible before they've even tried it, so the project stops before it even begins. :)

Cheers.

On Monday, March 13, 2017 at 11:57:49 AM UTC-7, Bryan Jones wrote:
I would imagine there is some risk of that, but I haven't seen that happen in my experiments. I carried out a negative control using just SLiCE extract with competent cells (No insert DNA), and didn't get any colonies on amp plates. Also the colonies I sequenced from my tests were all correct, no pKD46 carryover. It might be that DNAse breaks all the DNA from the SLiCE cells. Normally when you do a plasmid prep, you add EDTA to prevent this, but no EDTA was added here. I'm not positive why, but empirically, I can say that it does not seem to be an issue. 

On Tue, Mar 7, 2017 at 6:16 PM RBC <rco...@gmail.com> wrote:
Is there any worry that the amp resistant pKD46 plasmid will be extracted into the SLiCE lysate, in which case this plasmid will carry through to the final transformation? To me it would seem you can't use it to make amp resistant vectors..or maybe I'm missing something. Thanks for the clarification.

RC


On Friday, October 21, 2016 at 11:22:11 AM UTC-7, Bryan Jones wrote:
In my most recent test, without pKD46, I got 10 colonies from the SLiCE reaction, which was the same as the no SLiCE control. SLiCE with pKD46 resulted in 50-60 colonies. For comparison, NEBuilder resulted in about 100.



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[DIYbio] Human DNA sequencing

You can check out the 1000 genomes project, http://www.internationalgenome.org/ you can download genomic data from already assembled and annotated genomes all the way down to the raw sequencing data. I'm not sure if the metadata contains info on disease states of the inviduals, but you can always tie mutations back to known diseases.

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[DIYbio] Comment on APHIS regulatory proposals

Biohackers: it's comment on bad government GMO regulations time again. This time APHIS who is proposing to introduce process based regulation against the recommendations of the National Academy of Sciences. Please use this link to tell them this is a bad idea and will hurt innovation. Comments are due by Monday so let's get on this today.
https://www.regulations.gov/comment?D=APHIS-2015-0057-0001


Here's my full list of issues, more detail in the attached document. Feel free to use this to phrase your own comments.
1. It introduces process based regulation in violation of the 'product not process' principle of the coordinated framework and the recommendations of the National Academy of Sciences
2. This is a significant expansion of APHIS regulatory authority which is not justified by verifiable scientific risks, also in conflict with the principles of the coordinated framework. AHPIS own findings suggest GE plants pose no greater risk than conventionally bred plants
3. It inhibits innovation and disproportionally places the regulatory burden on small companies and academic researchers
4. The full negative impact of this clause has not been properly considered in the economic analysis of the rule
5. The breadth of the clause means many non-agricultural products will become regulated by APHIS which may be unintentional
6. No consideration has been made for grandfathering products which are currently legal to sell and distribute but which will become regulated under the new rules
7. The proposed weed risk assessment methodology is biased towards agricultural crops and is inadequate for the full range of plants which will be regulated by these proposals
8. Excessive regulation which is not aligned with actual risks could hurt USA competitiveness in the field of plant genetic engineering
9. The definition of Genetic Engineering proposed by AHPIS is not aligned with similar definitions being proposed by the FDA


thanks for your help!

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Re: [DIYbio] Human DNA sequencing

To sequence a human Genome, cheapest cost is about us $1000 per sample. You get a VCF file that lists all the mutations in that sample. You also get the raw data, about 100 g bytes per sample.

On Jun 16, 2017 4:37 AM, "Rajesh Solanki" <rbsolanki@gmail.com> wrote:
I would like to study disorders in human and relate with DNA.
I am thinking to get DNA sequenced fro few hundred persons.
I read few topic in group and it seems that building my own or buying DNA sequencer is not good idea.
What would be my options get it sequenced?
Also what are software option available to analyze the sequence data?

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[DIYbio] Human DNA sequencing

I would like to study disorders in human and relate with DNA.
I am thinking to get DNA sequenced fro few hundred persons.
I read few topic in group and it seems that building my own or buying DNA sequencer is not good idea.
What would be my options get it sequenced?
Also what are software option available to analyze the sequence data?

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Re: [DIYbio] Could we use optical heterodyning to boost UV bands into VIS?

On Wed, Jun 14, 2017 at 1:47 AM, John Ladasky <john.ladasky@gmail.com> wrote:
> I'm not sure exactly what this has to do with biology, but...

Mostly regarding the detection of biomolecules. I was thinking about
how to use cheaper light sources to substitute for less common
wavelengths, i.e. 260 and 280 nm.
The article I just posted is specifically about DNA research, and
implications of surface hydration effects on DNA interactions with
proteins, drugs, etc. It sounds pretty much like two-photon

> The standard 532 nm laser is actually a 1064 nm laser with a
> frequency-doubling crystal. Trying to make this relevant to biology,
> there's a technique called two-photon excitation microscopy

I'm familiar with both those concepts, I guess SFG is just a different
term for detecting the two-photon emissions that are characteristic of
certain molecules, which seem much weaker than the common two-photon
dyes. I wonder if it is a similar order of magnitude difference
between fluorescense and Raman scattering (reading now).

Very cool!

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[DIYbio] Fwd: CRISPR Collection: Editing the Genome




Annual Reviews: Food Allergy Collection
Read invited reviews written by world-renowned experts on the discovery and applications of CRISPR/Cas9 to cure human disease. These articles cover a variety of perspectives on genetic engineering platforms, animal models, and the potential advantages, drawbacks, and ethical considerations of CRISPR technology.

Access to this article collection is complimentary through July 31, 2017. Articles are always available for readers at subscribing institutions.

CRISPR/Cas9 pioneer Jennifer A. Doudna and Nobel Laureate Shinya Yamanaka are among the invited experts featured in this special collection of reviews covering advances in genetic engineering. Our articles are written by award-winning scientists and recognized leaders in their fields. Read selected articles now.

CRISPR-Cas9 Structures and Mechanisms
Fuguo Jiang and Jennifer A. Doudna, Annual Review of Biophysics

Evolution and Ecology of CRISPR
Edze R. Westra, Andrea J. Dowling, Jenny M. Broniewski, and Stineke van Houte
Annual Review of Ecology, Evolution, and Systematics

CRISPR/Cas9 in Genome Editing and Beyond
Haifeng Wang, Marie La Russa, and Lei S. Qi, Annual Review of Biochemistry

CRISPR/Cas9 for Human Genome Engineering and Disease Research
Xin Xiong, Meng Chen, Wendell A. Lim, Dehua Zhao, and Lei S. Qi
Annual Review of Genomics and Human Genetics

Engineering Delivery Vehicles for Genome Editing
Christopher E. Nelson and Charles A. Gersbach
Annual Review of Chemical and Biomolecular Engineering

Gene Editing: A New Tool for Viral Disease
Edward M. Kennedy and Bryan R. Cullen, Annual Review of Medicine

Genome Editing: A New Approach to Human Therapeutics
Matthew Porteus, Annual Review of Pharmacology and Toxicology

From Genomics to Gene Therapy: Induced Pluripotent Stem Cells Meet Genome Editing
Akitsu Hotta and Shinya Yamanaka, Annual Review of Genetics

We welcome you to share these articles with students and colleagues with an interest in research, policy, and patient care.
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