r******k
发帖数: 446 | 1 大神们推荐一下。 比如K562细胞 我怎么看它有什么gene被deletion或者amplication
？ 或者什么protein被磷酸化？？
多谢 |
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n******7
发帖数: 12463 | 2 对这个問題没经验
我感觉这种問題的一般思路就是评估一个background (H0)
只有严重偏离background的才是true signal
btw, 一般TFBS不是看调节区域吗，binding site在gene体的似乎不多？ |
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r**********e
发帖数: 587 | 3 或许我没说清楚。DE什么的我完全不懂，貌似是RNA-seq数据分析？differential
expression?（刚才查阅后才知道好像ChIP-seq也是可以用DESeq2来分析）
其实我真正研究的是某一种transposon element（只不过碰巧是某个TF ChIP-seq的
binding之一）。我其实做的事情就是根据repeatmasker，找出所有含有at least one
such transposon element的基因，把gene list直接放到GO term里去。
而我的背景list就是default的人类基因组所有的基因吧？
reference。
GO |
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N******n
发帖数: 3003 | 4 都看几个，找几个有意义的基因就行了。
channel gene 可以先排除，第二富集的的啥？ |
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发帖数: 1 | 6 大家好，
最近通过qpcr检测到几个基因的表达在我们的实验条件下有变化。所以我想把这个基因
克隆出来。但这个基因实在是没人研究(是个LINC)，所以没人有它的cDNA.所以我想试
试能否把它从cDNA克隆出来。我提取cDNA的办法是：
1. 用RNAeasy提取细胞的RNA,没有用shredder
2. 用iScript转录cDNA.引物用的就是它家的random primer
结果今天做了PCR,什么band都没有，那个郁闷。。。我克隆也做了几年了，很少失败过
。用的是NEB家的Q5.
想问一下大家，是我自己PCR条件没摸好，还是实验本身设计就行不通？我从来没做过
自己提取cDNA然后再克隆gene的。以前都是从公司买现成的cDNA. |
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z*******o
发帖数: 1794 | 7 对于没有人做过的基因我倒是觉得直接从CDNA里面克隆比较好，直接合成都是基于网上
预测，我还真碰到过自己克隆出来的序列和网上预测不一致的情况。好好纯化一下你的
RNA，多换几对引物，只要不是特别大的基因应该没有问题。
[在 imacoolboy (imacoolboy) 的大作中提到：]
:大家好，
:最近通过qpcr检测到几个基因的表达在我们的实验条件下有变化。所以我想把这个基
因克隆出来。但这个基因实在是没人研究(是个LINC)，所以没人有它的cDNA.所以我想试
:试能否把它从cDNA克隆出来。我提取cDNA的办法是：
:1. 用RNAeasy提取细胞的RNA,没有用shredder
:结果今天做了PCR,什么band都没有，那个郁闷。。。我克隆也做了几年了，很少失败
过。用的是NEB家的Q5.
:想问一下大家，是我自己PCR条件没摸好，还是实验本身设计就行不通？我从来没做过
:自己提取cDNA然后再克隆gene的。以前都是从公司买现成的cDNA. |
|
发帖数: 1 | 8 啊哈！我觉得就是这个！我用的是iscript cDNA first-strand kit, 它提供的是
oligodt 和 hexamer的mix.我明天就去订一个只有oligoDt的kit.
另外我想问一下，我有个gene它有6500bp左右长度。在做反转录的时候，您觉得反转的
时间大概需要多久。Biorad说在42℃上放60-90分钟。您觉得够嘛？
hexamer |
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k*****2
发帖数: 135 | 9 为什么不用gene-specific primer？小心二级结构复杂的时候或者有modification的时
候反转录酶不一定好用。你要不要试试ThermoScript™ RT-PCR System。可以打
电话让他们便宜点，我上次打电话给sales representative直接五折了。 |
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v********a
发帖数: 646 | 10 通过测序手段获得的一堆上调或者下降的基因(excel)
如何对其进行Gene Ontology/Function Annotation 分析
筛选到和该疾病相关的基因，从而进行下游分析？
没有编码经验，求指导，谢谢 |
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n***p
发帖数: 79 | 11 http://www.nature.com/news/ngago-gene-editing-controversy-escalates-in-peer-reviewed-papers-1.21023
摘录：
- Wensheng Wei, a molecular biologist at Peking University has already made
his mind up about NgAgo. “It simply doesn't work, period,” he says.
- Han told Nature he has discovered a problem that would not have been
obvious to others and that could explain why others are having difficulty
replicating his results. He says that he is currently running confirmatory
experiments so that he can publi... 阅读全帖 |
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发帖数: 1 | 12 想問一下你們gene synthesis 都用哪一家? |
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s********l
发帖数: 1195 | 13 Keesee K, Briggman JB, Thill G, Wu YJ (1996) Utilization of
nuclear matrix proteins for cancer diagnosis. CRC Crit Rev
Eukaryotic Gene Expr 6:189-214
Thanks! |
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m***n
发帖数: 266 | 16 听了几次关于这两大方面的seminar, 感觉很有意思。尤其是viral and non-viral gene
vector.
谁在做这方面的谈谈？ |
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b**s
发帖数: 589 | 17 use magnetic polymer?
gene |
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w*****q
发帖数: 2 | 18 做biosimilar的公司怎么知道一个蛋白的具体gene序列呢？专利上里有吗？如果有，那
做biosimilar的公司真是拣一大便宜啊。
查了几个，发现paper里的序列上都多少有点出入，到底以什么为准呢？ |
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b******1
发帖数: 1116 | 19 For Biosimilar, process is the prodcut... slightly different gene sequence
does not matter. |
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J****a
发帖数: 553 | 20 By my understanding, the DNA or amino acid sequence of the biosimilar
product need not be identical to the innovator product for a finding of
biosimilarity under the statute.There may be advantages to using a different
gene sequence that encodes the same protein.
"In general, FDA expects that the expression construct for a proposed
product will encode the same primary amino acid sequence as the reference
product. However, minor modifications such as N- or C-terminal truncations
that will not aff... 阅读全帖 |
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N******o
发帖数: 25 | 21 I guess it's OK to have different gene sequence, but final protein sequence
has to be identical, as FDA mentioned. Minor mofification generally referers
to post-translation modification, including glycosylation, C-terminal Lys
truncation for mAb. FDA does gives small rooms for those known modifications
, especially glycosylation. Those have to be justifitable based on bioassay,
preclinical, even clinical data.
Overall, I believe no sequence variation acceptable in term of biosimilar.
different
... 阅读全帖 |
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d********r
发帖数: 211 | 22 CMC development for cell & gene therapy
谁对这块熟悉? 可以给点信息吗?
多谢多谢!!! |
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c*****g
发帖数: 1856 | 23 我觉得clone gene don't need to cut the DNA off ba.
i think some ribozyme can work on this. |
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g******k
发帖数: 188 | 24
I don't know the correct answer either, that is why I am asking,
one of my committee member asked me about this
I think there are some tricky part, he mentioned about using lacZ opern
this is realy a tough question for those whose major in Molecular Biology.May
be only some guys from Harvard or MIT etc. could answer it,or it is very bad
question.
the question is"how would you clone a gene in 1950's, when the endonuclease
has not been dicovered yet?" |
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c*****g
发帖数: 1856 | 25 tho u can use mechnic method to break the DNA and then use the laz to pick
the gene fragment. maybe this can be done . still have many problem in d
detail. |
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c*****t
发帖数: 1879 | 26 I asked a microbiol professor. He gave me a possibility:
if you want to clone an antibiotic resistance gene, it can be
done by increasing the dosage of antibiotics, for instance, cam,
in order to clone (ie having more than 1 copy and on a plasmid).
Only cells w/ plasmids that can produce high copy can survive.
coco |
|
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d*******e
发帖数: 1649 | 28 define "gene expression problem" first |
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S******y
发帖数: 1123 | 29 Selecting subset of informative genes to predict binary outcome (such as
certain disease)... |
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s******g
发帖数: 6072 | 30 PhD第一年，正在考虑跟老板的问题，想问问各位大牛Stochastic modeling for gene
expression analysis这个方向如何？ |
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z****u
发帖数: 1007 | 31 不是做神经的，最近发现某个颌面部的干细胞分布和peripheral sensory nerve fiber
紧密相关。想找个inducible cre来在sensory nerve fiber里knockout掉一些gene.
请教该用啥cre比较好啊。最好是inducible cre. 多谢先了。 |
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z****u
发帖数: 1007 | 32 不是做神经的，最近发现某个颌面部的干细胞分布和peripheral sensory nerve fiber
紧密相关。想找个inducible cre来在sensory nerve fiber里knockout掉一些gene.
请教该用啥cre比较好啊。最好是inducible cre. 多谢先了。 |
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w*******y
发帖数: 60932 | 33 Star Trek The Animated Series - The Animated Adventures of Gene Roddenberry'
s Star Trek (1973)
Amazon Lightning Deal - - $13.99
VERY low price for this set. Buy quickly since when it's gone, it's gone!
linkola:
http://www.amazon.com/gp/product/B000HEWEJ4
|
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w*******y
发帖数: 60932 | 34 Link:
http://www.amazon.com/exec/obidos/ASIN/B004SH6XPM/ref=nosim/dea
Tax in KS, KY, ND, NY, WA.
This is the eBook version of the printed book.
This readable overview covers the rise of medical genetics through the past
century, and the eugenic impulses it has inspired. Nicholas Gillham reviews
the linkages between genes and disease; ethnic groups differential
susceptibility to genetic traits and disorders; personalized medicine; and
crucial social and ethical issues arising from the fields prog... 阅读全帖 |
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H******k
发帖数: 690 | 35 GENE最近跌了不少，能进点不？谁给说说？多谢了 |
|
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B*****e
发帖数: 2413 | 37 Since new patent policy will have more protections to genetics-related
research, all gene-related biotech company surge up today. |
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V*******s
发帖数: 19 | 38 Does anyone know how to analyze eukaryotic gene promoter sequence?
Thanks a lot. |
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G***G
发帖数: 16778 | 39 first you must answer why gene expression value vary from one experiment to
another experiment even in same condition and same tissue while sequences
keep unchanged.
from |
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s******9
发帖数: 283 | 40 NCBI protein database一般都有标注吧。
FEATURES Location/Qualifiers
source 1..503
/organism="Homo sapiens"
/db_xref="taxon:9606"
gene 1..503
/gene="TGFBR1"
/gene_synonym="ALK5"
/gene_synonym="SKR4"
Protein 1..503
/gene="TGFBR1"
/gene_synonym="ALK5"
/gene_synonym="SKR4"
... 阅读全帖 |
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k****s
发帖数: 1209 | 41 原文在
http://www.skizit.biz/2013/07/21/ecdysone-insect-hormone-an-ind
Ecdysone Insect Hormone Switches on Disease
21 Jul 2013 | ECDYSONE
MORGELLONS:
INSECT HORMONE ECDYSONE TRIGGERS GENETIC CHANGES
TO BUILD NEW HAIR, SKIN, NAILS and CAUSE DISEASES
SkizitGesture-2013
Morgellons is a military grade entomological terror weapon being used to
torture and kill innocent civilians. The medical community has not been
informed about this disease and its complexity keeps it from being properly
identified, ... 阅读全帖 |
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发帖数: 1 | 42
Download pdf published by SfN (Chapter from 'The History of Neuroscience in
Autobiography, Volume 8' edited by Larry R. Squire)
Yuh-Nung Jan's CV
Lily Jan's CV
Yuh-Nung Jan and Lily Jan
Birth
Family History and Growing Up
National Taiwan University
The Hiking Trip to Shitou in the Spring of 1967
Graduate School Application
Graduate Study at Caltech (1968锟974)
Seymour Benzer Lab (1974锟977)
Steve Kuffler锟絪 Lab at H... 阅读全帖 |
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n********k
发帖数: 2818 | 43 As you folks are discussing Cori, I happened to read his autobiography and
find it both interesting and enlightening:)))...BTW, it sounds Richard Axel
is a lot nicer than what I/we have heard...or in another word, his cruelty
is rooted in his obsession with science and indifference to humanity:))); As
well, it is pretty...he noted Cori but left his ex out...pretty human or
not human?
Autobiography
Richard AxelNew York City is my world. I was born in Brooklyn, the first
child of immigrant parents... 阅读全帖 |
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j*****8
发帖数: 12 | 44 I Have Made Significant and Internationally Recognized Scientific
Contributions to the Area of Medical Genetics, and to the Field of Genetics
of Mental Diseases in Particular
1. My significant contributions to XXXX at The University of Chicago (
Feb. 2005~Jun. 2008)
As Dr. GGGG stated, “It is well known that BBBBB disorder is one of
the most costly psychiatric conditions. It ranks among the top ten in
disability in the World Health Organization’s Global Burden of Disease.
More than 2... 阅读全帖 |
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P******l
发帖数: 1648 | 45 These are just google results from Apologetics websites for further study
and thoughts. Those who try to mess up with malicious attitude will get what
they deserve. On the other hand, we welcome sincere questions and are
willing to discuss about these together with mutual respect. Thanks.
http://askjohnmackay.com/blood-groups-how-a-b-ab-o-rh-blood-groups-from-adam-eve/
About the author: Diane Eager
Diane Eager has a background in medicine, and was a lecturer in biomedical
sciences at Univer... 阅读全帖 |
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A********e
发帖数: 354 | 46 【 以下文字转载自 Biology 讨论区 】
发信人: albertsmwk (.)(.), 信区: Biology
标 题: 第一批“青年千人计划”生物类@publication列表@
发信站: BBS 未名空间站 (Thu Aug 18 15:04:16 2011, 美东)
花了一个小时，深深的鄙视一下自己的无聊行径！！
125 蔡亮 男 1980年11月 复旦大学 生命
科学 2007年12月毕业于[美国]北卡大学 [美国]加州大学旧金山分校 博士后
Cai L, Mostov K. Polarity is destiny. Cell. 2009 Nov 13;139(4):660-2. PubMed
PMID: 19914162; PubMed Central PMCID: PMC2900917.
Cai L, Makhov AM, Schafer DA, Bear JE. Coronin 1B antagonizes cortactin and
remodels Arp2/3-containing actin branche... 阅读全帖 |
|
p*******g
发帖数: 2976 | 47 【 以下文字转载自 Biology 讨论区 】
发信人: albertsmwk (.)(.), 信区: Biology
标 题: 第一批“青年千人计划”生物类@publication列表@
发信站: BBS 未名空间站 (Thu Aug 18 15:04:16 2011, 美东)
花了一个小时，深深的鄙视一下自己的无聊行径！！
125 蔡亮 男 1980年11月 复旦大学 生命
科学 2007年12月毕业于[美国]北卡大学 [美国]加州大学旧金山分校 博士后
Cai L, Mostov K. Polarity is destiny. Cell. 2009 Nov 13;139(4):660-2. PubMed
PMID: 19914162; PubMed Central PMCID: PMC2900917.
Cai L, Makhov AM, Schafer DA, Bear JE. Coronin 1B antagonizes cortactin and
remodels Arp2/3-containing actin branche... 阅读全帖 |
|
O***n
发帖数: 13127 | 48 【 以下文字转载自 Biology 讨论区 】
发信人: albertsmwk (.)(.), 信区: Biology
标 题: 第一批“青年千人计划”生物类@publication列表@
发信站: BBS 未名空间站 (Thu Aug 18 15:04:16 2011, 美东)
花了一个小时，深深的鄙视一下自己的无聊行径！！
125 蔡亮 男 1980年11月 复旦大学 生命
科学 2007年12月毕业于[美国]北卡大学 [美国]加州大学旧金山分校 博士后
Cai L, Mostov K. Polarity is destiny. Cell. 2009 Nov 13;139(4):660-2. PubMed
PMID: 19914162; PubMed Central PMCID: PMC2900917.
Cai L, Makhov AM, Schafer DA, Bear JE. Coronin 1B antagonizes cortactin and
remodels Arp2/3-containing actin branche... 阅读全帖 |
|
a********k
发帖数: 2273 | 49 花了一个小时，深深的鄙视一下自己的无聊行径！！
125 蔡亮 男 1980年11月 复旦大学 生命
科学 2007年12月毕业于[美国]北卡大学 [美国]加州大学旧金山分校 博士后
Cai L, Mostov K. Polarity is destiny. Cell. 2009 Nov 13;139(4):660-2. PubMed
PMID: 19914162; PubMed Central PMCID: PMC2900917.
Cai L, Makhov AM, Schafer DA, Bear JE. Coronin 1B antagonizes cortactin and
remodels Arp2/3-containing actin branches in lamellipodia. Cell. 2008 Sep
5;134(5):828-42. PubMed PMID: 18775315; PubMed Central PMCID: PMC2570342.
Cai L, Makhov AM, Bear JE. F-actin... 阅读全帖 |
|
p*******g
发帖数: 2976 | 50 【 以下文字转载自 Biology 讨论区 】
发信人: albertsmwk (.)(.), 信区: Biology
标 题: 第一批“青年千人计划”生物类@publication列表@
发信站: BBS 未名空间站 (Thu Aug 18 15:04:16 2011, 美东)
花了一个小时，深深的鄙视一下自己的无聊行径！！
125 蔡亮 男 1980年11月 复旦大学 生命
科学 2007年12月毕业于[美国]北卡大学 [美国]加州大学旧金山分校 博士后
Cai L, Mostov K. Polarity is destiny. Cell. 2009 Nov 13;139(4):660-2. PubMed
PMID: 19914162; PubMed Central PMCID: PMC2900917.
Cai L, Makhov AM, Schafer DA, Bear JE. Coronin 1B antagonizes cortactin and
remodels Arp2/3-containing actin branche... 阅读全帖 |
|
