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Fwd: bionet.molbio.methds-reagnts - 2 new messages in 2 topics - digest
From: "bionet.molbio.methds-reagnts group" <noreply@googlegroups.com>
Date: Dec 28, 2010 3:00 AM
Subject: bionet.molbio.methds-reagnts - 2 new messages in 2 topics - digest
To: "bionet.molbio.methds-reagnts digest subscribers" <bionet.molbio.methds-reagnts@googlegroups.com>
bionet.molbio.methds-reagnts
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Today's topics:
* Methods Digest, Vol 67, Issue 8 - 1 messages, 1 author
http://groups.google.com/group/bionet.molbio.methds-reagnts/t/ca739b039a1c5a1f?hl=en
* gel elution of 6KB PCR product - 1 messages, 1 author
http://groups.google.com/group/bionet.molbio.methds-reagnts/t/fcdaf69d6c779d50?hl=en
==============================================================================
TOPIC: Methods Digest, Vol 67, Issue 8
http://groups.google.com/group/bionet.molbio.methds-reagnts/t/ca739b039a1c5a1f?hl=en
==============================================================================
== 1 of 1 ==
Date: Sun, Dec 26 2010 5:35 am
From: Virash Gupta
Dear B Ram,
try following following method. make a very small hole at the bottom
of a 0.5 ml PCR tube and cover it with some sterile glass wool. Put
your cut gel band in this tube and close it. Now put this tube into a
1.5 ml eppendorf tube. using an appropriate balancing tube spin at
12,000 rpm for 5 min. you should get 70-80% of your DNA in the
eppendorf tube which can be precipitated and resuspended in small
volume. Alternatively, put this tube into 1.5 ml tube containing ~100
microlitre TAE buffer. Also add same amount of this buffer in 0.5 ml
tube. Take two small pieces of platinum wires, dip one through space
between 0.5 ml and 1.5 ml tube so that it immerses in the buffer.
other piece of platinum wire should be purt in 0.5 ml tube buffer.
apply low current (~ 10V) with black electrode connected to wire in
0.5 ml tube and red electrode to other one immersed in 1.5 ml tube
buffer. This will make a small electrophoresis coloumn. after around
20 min, whole of DNA will move into buffer of 1.5 ml tube. Disacrd 0.5
ml tube along with its contents. precipitate DNA from buffer of 1.5 ml
tube using sod acetate and cold ethanol. All the best.
On 12/25/10, methods-request@oat.bio.indiana.edu
<methods-request@oat.bio.indiana.edu> wrote:
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> 1. gel elution of 6KB PCR product (B.Rama chandran)
>
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> ----------------------------------------------------------------------
>
> Message: 1
> Date: Sat, 25 Dec 2010 18:52:35 +0530
> From: "B.Rama chandran" <chandranbrama@gmail.com>
> Subject: gel elution of 6KB PCR product
> To: Methods@magpie.bio.indiana.edu
> Message-ID:
> <AANLkTikscKjcve5M+V_SpxhzymrfjbiQ39HRep0YQOCS@mail.gmail.com>
> Content-Type: text/plain; charset=UTF-8
>
> Dear All,
>
> I am facing problem in the gel elution of 6KB PCR product, I am
> getting very less yield (~10ng/micto liter). I am using Sigma gel
> extraction kit (catalog no:NA1111). Since yield is very less I couldn't use
> that product for restriction digestion. If you have any suggestion please
> give me. If you know any other technique which will give better yield please
> let me know that.
>
> regards,
> B.Ram.
>
>
> ------------------------------
>
> _______________________________________________
> Methods mailing list
> Methods@net.bio.net
> http://www.bio.net/biomail/listinfo/methods
>
> End of Methods Digest, Vol 67, Issue 8
> **************************************
>
--
Dr V K Gupta
Sr Microbiologist (Mol Biology)
IMBL, Department of Entomology
Pun. Agric. Univ., Ludhiana (Pb)-141004- India
M: 081465-55515
==============================================================================
TOPIC: gel elution of 6KB PCR product
http://groups.google.com/group/bionet.molbio.methds-reagnts/t/fcdaf69d6c779d50?hl=en
==============================================================================
== 1 of 1 ==
Date: Mon, Dec 27 2010 2:12 am
From: Fathi Hassan
hello,
you can try to clone the PCR product you got from gel in any cloning vector like
pJet or pGem, since they require low amounts of PCR products, then you can
digest the vector and get the fragment ready for further cloning. it is two
steps more but may help you
good luck
FH
________________________________
From: DK <dk@no.email.thankstospam.net>
To: methods@magpie.bio.indiana.edu
Sent: Sun, December 26, 2010 12:28:15 AM
Subject: Re: gel elution of 6KB PCR product
In article <mailman.445.1293295295.15153.methods@net.bio.net>, "B.Rama chandran"
<chandranbrama@gmail.com> wrote:
>Dear All,
>
> I am facing problem in the gel elution of 6KB PCR product, I am
>getting very less yield (~10ng/micto liter). I am using Sigma gel
>extraction kit (catalog no:NA1111). Since yield is very less I couldn't use
>that product for restriction digestion. If you have any suggestion please
>give me. If you know any other technique which will give better yield please
>let me know that.
What is the yield as % of input? If you didn't have much to begin
with, nothing can help. ~50% on gel-extraction are ~ normal. If it is the
yield that is poor, try the same protocol with 2 kbp. If the result is good,
your kit is not appropriate for 6K. If the result is bad, either your
reagents are gone bad/improperly prepared or you are doing something
wrong. I've purified 14 kbp from gel with about 30% using Qiagen's
kit. Never a problem.
DK
_______________________________________________
Methods mailing list
Methods@net.bio.net
http://www.bio.net/biomail/listinfo/methods
--0-1362310605-1293367365=:9174
Content-Type: text/html; charset=us-ascii
<html><head><style type="text/css"><!-- DIV {margin:0px;}
--></style></head><body><div
style="font-family:arial,helvetica,sans-serif;font-size:12pt">you can try to
clone the PCR product you got from gel in any cloning vector like pJet or pGem,
since they requier low amounts of PCR products, then you can digest the vector
and get the fragment ready for further cloning. it is two steps more but may
help you<br>good luck<br>FH<br><br><div style="font-family:
arial,helvetica,sans-serif; font-size: 12pt;"><div style="font-family:
arial,helvetica,sans-serif; font-size: 12pt;"><font face="Tahoma" size="2"><hr
size="1"><b><span style="font-weight: bold;">From:</span></b> DK
<dk@no.email.thankstospam.net><br><b><span style="font-weight:
bold;">To:</span></b> methods@magpie.bio.indiana.edu<br><b><span
style="font-weight: bold;">Sent:</span></b> Sun, December 26, 2010 12:28:15
AM<br><b><span style="font-weight: bold;">Subject:</span></b> Re: gel elution of
6KB PCR product<br></font><br>
In article <<a
ymailto="mailto:mailman.445.1293295295.15153.methods@net.bio.net"
href="mailto:mailman.445.1293295295.15153.methods@net.bio.net">mailman.445.1293295295.15153.methods@net.bio.net</a>>,
"B.Rama chandran" <<a ymailto="mailto:chandranbrama@gmail.com"
href="mailto:chandranbrama@gmail.com">chandranbrama@gmail.com</a>>
wrote:<br>>Dear All,<br>><br>> I am facing problem
in the gel elution of 6KB PCR product, I am<br>>getting very less yield
(~10ng/micto liter). I am using Sigma gel<br>>extraction kit (catalog
no:NA1111). Since yield is very less I couldn't use<br>>that product for
restriction digestion. If you have any suggestion please<br>>give me. If you
know any other technique which will give better yield please<br>>let me know
that.<br><br>What is the yield as % of input? If you didn't have much to begin
<br>with, nothing can help. ~50% on gel-extraction are ~ normal. If it
is the <br>yield that is poor, try the same protocol with 2 kbp. If the result
is good, <br>your kit is not appropriate for 6K. If the result is bad, either
your <br>reagents are gone bad/improperly prepared or you are doing something
<br>wrong. I've purified 14 kbp from gel with about 30% using Qiagen's <br>kit.
Never a problem.
<br><br>DK<br>_______________________________________________<br>Methods mailing
list<br><a ymailto="mailto:Methods@net.bio.net"
href="mailto:Methods@net.bio.net">Methods@net.bio.net</a><br><span><a
target="_blank"
href="http://www.bio.net/biomail/listinfo/methods">http://www.bio.net/biomail/listinfo/methods</a></span><br></div></div>
</div><br>
</body></html>
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Monday, December 27, 2010
Sunday, December 26, 2010
ubuntu on Android
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single crystal diamond NEMS switch
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microscope on a chip
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patterning of viruses by dip pen nanolithography
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spintronic computer
Fwd: The 10 insights of the decade, 10 breakthroughs of 2010 (K21st - Essential 21st Century Knowledge)
From: "Newsfeed to Email Gateway" <emlynoregan@gmail.com>
Date: Dec 17, 2010 11:46 AM
Subject: The 10 insights of the decade, 10 breakthroughs of 2010 (K21st - Essential 21st Century Knowledge)
To: <technologiclee@gmail.com>
The 10 insights of the decade, 10 breakthroughs of 2010 (12/17/10 14:50:45 UTC)
From quantum mechanics, unveiling and sequencing the Neandertal genome, to Exoplanets and metamaterials, what more could we ask for?
"This year's Breakthrough of the Year represents the first time that scientists have demonstrated quantum effects in the motion of a human-made object," said Adrian Cho, a news writer for Science. "On a conceptual level that's cool because it extends quantum mechanics into a whole new realm. On a practical level, it opens up a variety of possibilities ranging from new experiments that meld quantum control over light, electrical currents and motion to, perhaps someday, tests of the bounds of quantum mechanics and our sense of reality."
Science's list of the nine other groundbreaking achievements from 2010 follows.
Synthetic Biology: In a defining moment for biology and biotechnology, researchers built a synthetic genome and used it to transform the identity of a bacterium. The genome replaced the bacterium's DNA so that it produced a new set of proteins—an achievement that prompted a Congressional hearing on synthetic biology. In the future, researchers envision synthetic genomes that are custom-built to generate biofuels, pharmaceuticals or other useful chemicals.
Neandertal Genome: Researchers sequenced the Neandertal genome from the bones of three female Neandertals who lived in Croatia sometime between 38,000 and 44,000 years ago. New methods of sequencing degraded fragments of DNA allowed scientists to make the first direct comparisons between the modern human genome and that of our Neandertal ancestors.
HIV Prophylaxis: Two HIV prevention trials of different, novel strategies reported unequivocal success: A vaginal gel that contains the anti-HIV drug tenofovir reduced HIV infections in women by 39 percent and an oral pre-exposure prophylaxis led to 43.8 fewer HIV infections in a group of men and transgender women who have sex with men.
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Quantum Simulator: To describe what they see in the lab, physicists cook up theories based on equations. Those equations can be fiendishly hard to solve. This year, though, researchers found a short-cut by making quantum simulators—artificial crystals in which spots of laser light play the role of ions and atoms trapped in the light stand in for electrons. The devices provide quick answers to theoretical problems in condensed matter physics and they might eventually help solve mysteries such as superconductivity.
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The Return of the Rat: Mice rule the world of laboratory animals, but for many purposes researchers would rather use rats. Rats are easier to work with and anatomically more similar to human beings; their big drawback is that methods used to make "knockout mice"—animals tailored for research by having specific genes precisely disabled—don't work for rats. A flurry of research this year, however, promises to bring "knockout rats" to labs in a big way.
Finally, to celebrate the end of the current decade, Science news reporters and editors have taken a step back from their weekly reporting to take a broader look at 10 of the scientific insights that have changed the face of science since the dawn of the new millennium. A list of these 10 "Insights of the Decade" follows.
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Water on Mars: Half a dozen missions to Mars over the past decade have provided clear evidence that the Red Planet once harbored enough water—either on it or just inside it—to alter rock formations and, possibly, sustain life. This Martian water was probably present around the time that life was beginning to appear on Earth, but there is still enough moisture on Mars today to encourage scientists seeking living, breathing microbes.
Reprogramming Cells: During the past decade, the notion that development is a one-way street has been turned on its head. Now, researchers have figured out how to "reprogram" fully developed cells into so-called pluripotent cells that regain their potential to become any type of cell in the body. This technique has already been used to make cell lines from patients with rare diseases, but ultimately, scientists hope to grow genetically matched replacement cells, tissues and organs.
The Microbiome: A major shift in the way we view the microbes and viruses that call the human body home has led researchers to the concept of the microbiome—or the collective genomes of the host and the other creatures that live on or inside it. Since 90 percent of the cells in our bodies are actually microbial, scientists are beginning to understand how significantly microbial genes can affect how much energy we absorb from our foods and how our immune systems respond to infections.
Exoplanets: In the year 2000, researchers were aware of just 26 planets outside our solar system. By 2010, that number had jumped to 502—and still counting. With emerging technologies, astronomers expect to find abundant Earth-like planets in the universe. But for now, the sizes and orbits of larger planets already discovered are revolutionizing scientists' understanding of how planetary systems form and evolve.
Inflammation: Not long ago, inflammation was known as the simple sidekick to our healing machinery, briefly setting in to help immune cells rebuild tissue damage caused by trauma or infection. Today, however, researchers believe that inflammation is also a driving force behind the chronic diseases that will eventually kill nearly all of us, including cancer, Alzheimer's disease, atherosclerosis, diabetes and obesity.
Metamaterials: By synthesizing materials with unconventional and tunable optical properties, physicists and engineers have pioneered new ways to guide and manipulate light, creating lenses that defy the fundamental limits on resolution. They've even begun constructing "cloaks" that can make an object invisible.
Climate Change: Over the past decade, researchers have solidified some fundamental facts surrounding global climate change: The world is warming, humans are behind the warming and the natural processes of the Earth are not likely to slow that warming. But, the next 10 years will determine how scientists and policymakers proceed with this vital information.
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microscope moves atoms
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Android 2.3 Platform | Android Developers
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Fwd: Rev
From: Lee Nelson <technologiclee@gmail.com>
Date: Mon, Jun 14, 2010 at 1:02 PM
Subject: Rev
To: Lee Nelson <technologiclee@gmail.com>
Title:ct cs
Sunday, December 5, 2010
Nanoengineers wiki
Welcome to your new Wikidot site - Nanoengineers
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Subject: Google Alert - "xray laser"
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Friday, December 3, 2010
OPTICAL TRAP BONDING
SYSTEM AND METHOD FOR HOLOGRAPHIC OPTICAL TRAP BONDING - Patent application
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Thursday, December 2, 2010
Fwd: comp.lang.python - Wing IDE
Hi,
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