c**i
发帖数: 6973 | 1 A Taiwanese businessman and his associates tried to steal the method to
chemically synthesize 紫杉醇, got caught and went to US prison. |
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a****d
发帖数: 1919 | 2 Two step PCR should work for your propose.
synthesize two pair of primers:
F1-xxxx
30bp-xxxx-R1
F2-xxx-30bp
xxxx-R2
PCR separately, then put together both products as template, PCR again using
F1-xxxx and xxx-R2. The final PCR product could be subcloned into the
plasmid. You have to find the suitable digestion sites when designing
primers though. |
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l****z
发帖数: 29846 | 3 好像图转不过来. 这是link:
http://snowball5212.blog.hexun.com/70784459_d.html
中国人一直自称是中国人首次人工合成了胰岛素, 也是首次人工合成蛋白质, 并为这一
成果没有获得诺贝尔奖而不平, 时常有人说“诺贝尔奖是西方人的奖”时就会提起这茬
。中国人率先人工合成蛋白质, 这确实是官方说法, 并出现在中小学历史教材里, 作为
共和国科技发展迅猛的案例。
具体报告时间可以在上图中看到, 以上两图片来自《结晶胰岛素的全合成》一书。
但是, 国际上根本没有承认这一“伟大成就”的“伟大”, 甚至归功他人。在THE
HISTORY OF INSULIN 一文里, 提到“In fact, insulin was the first protein to
be chemically synthesized in a laboratory, in 1963. ”，你会注意到, 这比我国
所说要早了两年。在《Insulin & related proteins: structure to function and
pharmacology》一书... 阅读全帖 |
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b******y
发帖数: 627 | 4 You can custom synthesize the whole thing with a few hundred dollars. |
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o**4
发帖数: 35028 | 5 How much to synthesize 5kb DNA? |
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c********b
发帖数: 363 | 6 I just synthesized one by myself. Mine is 408 bp. I ordered 8 primers (
contains short complementary sequence one after another). The first 4 (
forward, reverse, forward, reverse to cover the first half of the sequence)
primers and the last 4 primers were put into 2 PCRs for annealing and
filling. The PCR products were then put into a new PCR for annealing and
filling. Then used the first forward primer and the last reverse primer for
a regular PCR. Gel purified the product, cloning. One day's j... 阅读全帖 |
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e**e
发帖数: 166 | 7 Glycobiology vol. 22 no. 3 pp. 439–451, 2012
The ability of land plants to synthesize glucuronoxylans
predates the evolution of tracheophytes
Can I have the supplement data please?
please send to e****[email protected]
Thank you. |
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n*****k
发帖数: 69 | 8 Thanks a lot!
By the way, how to get the "Smac mimetic"? commercial available? or need to
synthesize? |
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g*********5
发帖数: 2533 | 9 if I have a peptides sequence,
you help me synthesize the peptide and link it to KLH, inject rabbits and
10week service. how much you charge?
thanks |
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h**********r
发帖数: 671 | 10 We even don't have money to synthesize the gene.
So are there any labs of 老板是炸药奖或被提名过的 poor like this? |
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s******9
发帖数: 283 | 11 total mRNA for cell-free translation is a bad idea, unless you have single-
molecule detection sensitivity. You have to fish out the cDNA, add a strong
promoter and synthesize a large amount of mRNAs for the translation if you
can only do western. |
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b******y
发帖数: 627 | 12 Short peptides may have low immunogenecity. To increase the chance of
getting Ab, you can synthesize a peptide dendrimer from C TERMINUS to N
terminus (as all peptide synthesis is from C to N): as following:
EK(K(K(linker-your peptide)2)2)2.
In this way, you will have a dendrimer that have 2^n of your peptides
presented outwards. This kind of dendrimer has increased immunogenecity and
therefore higher chance to generate Ab.
Anaspec should have this service although a bit expensive.
good luck. |
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b******y
发帖数: 627 | 13 It will be expensive from Anaspec. should try luck with smaller company for
this.
Do you have friends who can help you with this? It is not different from
normal peptide synthesis except that the protection on the two -NH2 on the
first Lysines are the same probably fmoc. You also don't mind a mixture.
There is one more strategy if your peptide doesn't have cys in it.
Synthesize your peptide with an extra cys at the C-terminus. Buy tri-
maleimide from somewhere. Mix your peptide with tri-maleimid... 阅读全帖 |
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b******y
发帖数: 627 | 14 Not complicated for the last strategy. free thiol from cys reacts with
maleimide very efficiently. tri-maleimide is cheap. you have to synthesize
peptide anyway.
Just a reminder: the N-termini of proteins in vivo (bacteria or mammalian
cell) are complicated. The majority of them have the met cleaved upon
translation. Also the majority of them are N-terminally acetylated
cotranslationally or post-translationally.
Be careful! Just want to remind you not to use wrong peptides for Ab
generation. |
|
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w***7
发帖数: 1637 | 16 source：http://bbs.netbig.com/thread-2491712-1-2.html
看了一下，今年不错啊，和清华并驾齐驱。
而且下半年我知道的还有两篇NS。
~~~~~~~~~~~~~~~~~
1.
Yuan W, Wu T, Fu H, Dai C, Wu H, Liu N, Li X, Xu M, Zhang Z, Niu T, Han Z,
Chai J, Zhou XJ, Gao S, Zhu B
Dense Chromatin Activates Polycomb Repressive Complex 2 to Regulate H3
Lysine 27 Methylation
Science, DOI: 10.1126/science.1225237, Aug. 24, 2012
2.
Liu B, Du H, Rutkowski R, Gartner A, Wang X
LAAT-1 Is the Lysosomal Lysine/Arginine Transporter That Maintains Amino
Acid Homeostasis
S... 阅读全帖 |
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l**********1
发帖数: 5204 | 17 Agree with sunny said.
btw, for 肽类analog from mushrom
pls refer
one 2011 review:
by
Wasser SP (2011)
Current findings, future trends, and unsolved problems in studies of
medicinal mushrooms.
Appl Microbiol Biotechnol. 89: 1323-32.
Its PubMed link:
http://www.ncbi.nlm.nih.gov/pubmed/21190105
and for 肽类analog modification to anti-tumor drugs
pls refer,
by
Iwasaki T et al. (2009)
Selective cancer cell cytotoxicity of enantiomeric 9-mer peptides derived
from beetle defensins depends on negatively ... 阅读全帖 |
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A******y
发帖数: 2041 | 18 Na, nucleotides synthesized in solution phase could be very cheap, too. The
cost of the drug is never an issue. There are recombinant protein
therapies that cost more than 300k a year and medicare/medicaid pays for
them.
The delivery just too difficult. Nature over billions of years evolution
only given virus as only effective way to deliver nucleotides. I think we
are still not at the point of out-smart nature. Also, the idea of magic
bullet is fading, you need a magic shotgun that can hit... 阅读全帖 |
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b******n
发帖数: 4225 | 19 1 Science 2 Cell papers:
Devireddy LR, Teodoro JG, Richard FA, Green MR (2001) Induction of
apoptosis by a secreted lipocalin that is transcriptionally regulated by IL-
3deprivation. Science 293: 829–834.
Devireddy LR, Gazin C, Zhu X, Green MR (2005) A cell-surface receptor for
lipocalin 24p3 selectively mediates apoptosis and iron uptake. Cell 123:
1293–1305.
Devireddy, L. R., D. O. Hart, D. H. Goetz & M. R. Green, (2010) A mammalian
siderophore synthesized by an enzyme with a bacterial homolog... 阅读全帖 |
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b******n
发帖数: 4225 | 20 1 Science 2 Cell papers:
Devireddy LR, Teodoro JG, Richard FA, Green MR (2001) Induction of
apoptosis by a secreted lipocalin that is transcriptionally regulated by IL-
3deprivation. Science 293: 829–834.
Devireddy LR, Gazin C, Zhu X, Green MR (2005) A cell-surface receptor for
lipocalin 24p3 selectively mediates apoptosis and iron uptake. Cell 123:
1293–1305.
Devireddy, L. R., D. O. Hart, D. H. Goetz & M. R. Green, (2010) A mammalian
siderophore synthesized by an enzyme with a bacterial homolog... 阅读全帖 |
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n***3
发帖数: 663 | 21 Nat Biotechnol. 2012 Nov 25. doi: 10.1038/nbt.2436.
Protective efficacy of in vitro synthesized, specific mRNA vaccines against
influenza A virus infection.
Please send the paper to e*********[email protected]
Thanks! |
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G****a
发帖数: 10208 | 22 【 以下文字转载自 Missouri 讨论区 】
发信人: Geisha (朔风如解意 容易莫摧残), 信区: Missouri
标 题: 钱mm的文章都是ERIC的， ERIC让她当第一作者
发信站: BBS 未名空间站 (Sun Jan 27 19:16:10 2013, 美东)
http://www.qianlab.caltech.edu/Qian_CV.pdf
Refereed Publications (* co-first authors)
1. L. Qian, E. Winfree, and J. Bruck. Neural network computation with DNA
strand displacement cascades. Nature, 475:368-372, 2011.
News and Views: “DNA and the brain” by Anne Condon, Nature, 475:304-305.
Media coverage: msnbc News, CBS News, Discovery News, CNET News, D... 阅读全帖 |
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Z*****1
发帖数: 20 | 23 我在化学实验室，现在正在合成一个15mer peptide， 但是仪器一直有问题。 具体表
现就是在coupling那一步，溶液应该是从amino acid vial 转移到 reaction vessel
然后 coupling， 但是溶液被直接转移到waste container。所以，reaction vessel
里没有任何溶液，也就没法继续了。 老板不想花钱请人过来修，我也没办法。不知道
有没有人遇到过同样的情况？先谢过了…… |
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f****h
发帖数: 20 | 24 试着直接给公司打电话问问吧。我们的PS3出过两次问题，一次是感应AA vial的传感器
坏了，在保修期内，公司直接派人来修。还有一次carousel总是错位，给公司打电话，
tech遥控指挥我们自己修好的 |
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s******9
发帖数: 283 | 25 人说：
"If this takes away from any of the R01s that would normally be funded by
the NIH, it would be bad. Right now the community is already so strapped we'
re at a breaking point."
called several of its technological proposals "science fiction"
...
http://www.sciencemag.org/content/339/6123/1022.full
Brain Project Draws Presidential Interest, but Mixed Reactions
Emily Underwood*
Shortly after President Barack Obama made a seemingly innocuous pitch for
more brain research in his State of the Union... 阅读全帖 |
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k****l
发帖数: 279 | 26 If you synthesize that DNA, just add a dideoxy C in the 3' end |
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H*****e
发帖数: 120 | 27 Here are some information that may be interesting to you. But you need to
check the literature yourself since I heard most of them from a friend
working on it.
1. Acetate-T is the synthesized form that chemist believe will be
bioavailable. If you can find some PK/PD data, it will be believable. Succi
-T is the so-called conjugated form in vivo, a product by Phase I and II
enzymes. Many chemicals will be converted to this kind of form in liver to
further process. I am not sure what kind of ex... 阅读全帖 |
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s**********e
发帖数: 2888 | 28 found an interesting article:
One of the favourite arguments of the pro-GM lobby in support of Bt-toxin GM
crops is that the Bt toxin has been safely used for decades by organic
farmers.
Many thanks to Susan Pusztai for the following information which precisely
delineates the very considerable differences between the use of Bt in
organic farming and the use of the Bt toxin as a transgene in GM crops.
This should be read in conjunction with Arpad Pusztai's recent comments,
posted to the ngin list... 阅读全帖 |
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L*********y
发帖数: 322 | 29
You might want to do more function-structural analysis of this transporter
to back up your data.
For example, within this transporter, can you identify the key amino acid
residues or protein domains that affect the compound binding? Second,
can you synthesize a fluorophore or radiation conjugated version of this
new compound you discovered? If yes, you should be able to test its
interaction with your transporter by IP.
Third, Can you do some in vivo experiments to support you in vitro findings... 阅读全帖 |
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q******g
发帖数: 3858 | 30 For example, within this transporter, can you identify the key amino acid
residues or protein domains that affect the compound binding?
We will examine the amino acid uptake in oocytes expressing mutant with
compound.
Second, can you synthesize a fluorophore or radiation conjugated version of
this new compound you discovered? If yes, you should be able to test its
interaction with your transporter by IP.
This is a revesible inhibitor for transporter. I am afraid that compound
will not bind to t... 阅读全帖 |
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g*********3
发帖数: 177 | 31 Just my opinion/impression:
1) How to define "TCA is inhibited":
Sometimes it is not totally inhibited in that whole TCA Cycle. It has
another name: Truncated TCA cycle.
2) as xhzhou said: the intermediate products in glycolysis can enter into
other pathways---"whatever pathways"---synthesized into "macromolecules".
The "whatever pathways" & "macromolecules" are hot filed, which is more
complex than the textbook. Many of them are not discovered yet and may be
tumor-specific SUCH as the oncometab... 阅读全帖 |
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g*********5
发帖数: 2533 | 32 IDT could synthesize 3000bp now. |
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H****N
发帖数: 997 | 33 Anybody have experience of synthesizing large (~10 kb) DNA fragments? Which
company do you use? Thank you. |
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H****N
发帖数: 997 | 34 Anybody have experience of synthesizing large (~10 kb) DNA fragments? Which
company do you use? Thank you. |
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r*******y
发帖数: 48 | 35 you just need synthesized ssODN as a template for transfection; it is not
necessary to subclone it into plasmd and then release it. |
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H****S
发帖数: 40 | 36 Just synthesize ss donor with G-block @ IDTdna
2-3kb is easy for G-block synthesis |
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s****l
发帖数: 10462 | 37 有人愿意讨论这里面提及的测序技术吗？优缺点，可行性，第一手经验等等
这里面谁现在是赢家很清楚，但是可见的将来和更远的将来谁会赢？
http://cen.acs.org/articles/92/i33/Next-Gen-Sequencing-Numbers-
全文拷贝如下
试试看图能不能贴上
/1407973177171.jpg
/1408028636124.jpg
OVER STORY
Next-Gen Sequencing Is A Numbers Game
As technical and cost barriers fall, instrument firms move their systems
into research and clinical markets
By Ann M. Thayer
Department: Business
Keywords: instrumentation, gene sequencing, diagnostics, genomics, cancer
[+]Enlarge
09233-cover-Openercxd_17647969-690
A... 阅读全帖 |
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z******n
发帖数: 97 | 38 The most efficient way to identify a hit is to look at ligands biding to
similar proteins if no existing hits available, develop a robust assay and
design/synthesize some similar analogs to test, or buy a library which is
most likely very expensive. If you are lucky, you may end up with weak but
solid hits, however, the danger is that many molecules reported by academic
groups are covalently reactive, which may give false hit, and there is a
Nature review talking about this. Regarding computer d... 阅读全帖 |
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w******f
发帖数: 37 | 39 谢谢！
另外， 我打算synthesize loxp-STOP-loxp cassette，但苦于找不到已经证明可行的
sequence（希望STOP是比较短的sequence）。有可推荐的么？谢谢 |
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I****7
发帖数: 6 | 40 组里面要在4-5月份招一个多肽合成，本科或者硕士。不解决身份问题，需要有过多肽
合成经验，会熟练操作hplc, mass spec, synthesizer。请信内联系并留电话。 |
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s******y
发帖数: 28562 | 44 对。
如果是混在一起的普通Mass Spec 不行的，因为对方公司会耍赖说普通质谱没有定量性。 |
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K******S
发帖数: 10109 | 45 triple A analysis, all synthetic peptides should have AAA info. |
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v**********t
发帖数: 9 | 46 这是他们给的图谱，纯吗？
各位懂得给看看。
谢谢。 |
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C******T
发帖数: 300 | 47 LCMS 和HPLC 都只能大致判断peptide的纯度，无法定量。
这两种方法（LC和HPLC）是依靠紫外峰估算peptide的含量。很多时候样品里的salt
and small molecule impurity不能依靠紫外吸收分析。业界认可的定量方法是楼上说
的AAA analysis。所谓 75%， 90% peptide content, 指的是AAA analysis 得到的数
据。
一个peptide经过HPLC提纯，看似纯度达到95%，做一下AAA analysis，发现peptide
content 只有50%，是很常见的事情。
不过楼上的样品，根据MS谱图来看，没有做AAA analysis的价值，太不纯了。50AA左右
的多肽，应该只有四五个峰。机器灵敏度不高的话，一般就是两三个峰。 |
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K******S
发帖数: 10109 | 49 It's pretty standard procedure. It more depends on your peptide's sequence. |
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v**********m
发帖数: 5516 | 50 公司合成时会出specification，AAA通常是做过的，会给出(weight of多肽/weight of
样品)的纯度。那些一起干燥出来的小分子如醋酸盐等都会加重样品的表观重量。这个
只会对绝对浓度有影响,另外小分子对后续的生物实验影响不大，透析一下就可去除。
我觉得LZ应该是更关心多肽内部的纯度，合成步骤多时肯定有一些会少掉某些AA，那些
多肽杂质如果太多，会影响后续的生物研究。此类杂质只能用RP-HPLC鉴定，LCMS能更
一步的定性多肽杂质。这些多肽杂质和目的多肽性质理化接近，只能过制备柱进一步纯
化。
LZ的质谱图貌似MALDI-TOF，这个质谱样品的matrix非常杂，有些杂质也许是样品本身
带的，也可能是学校的facility质谱机器太脏导致的。如果能走LC-ESI-QTOF或TOF，基
本能确定多肽杂质在总多肽里所占的比例。 |
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