c********n
发帖数: 225 | 1 谈谈和蛋白打交道的日子 （一）
序
笔者已经做了多年和蛋白相关的科研与服务，产品研发生产以及产业化。在这里分享一
下与蛋白打交道这些年带来的一些感想。其实所有的科学研究都是用来解决人类已经和
即将遇到的各种各样的问题的，只有解决了问题才会有更大的存在价值。
第一篇 DNA测序， Sanger Style - 纪念Frederick Sanger
2013年11月20号，Frederick Sanger 去世了， 享年95岁。Sanger是DNA测序之父，获
得过两次诺贝尔奖， 一个是1958年蛋白测序， 另一个是1980年DNA测序。虽然DNA结构
发现的比蛋白晚， 但毕竟是genetic code，master script。ATCG四个碱基的组合比蛋
白的20个氨基酸的天文数字组合可更容易研究找规律了。人天生就是个好奇的动物，
最喜欢找规律, 尤其喜欢从简单的地方入手，所以围绕DNA的研究在Watson-Crick的碱
基配对这个简单地规律被发现以后，起来的就比蛋白快了很多。其实DNA和蛋白， 就像
是帅才和将才， 他们的用处是不同的，需要出色的帅才（DNA）订好了方向，能干的将... 阅读全帖 |
|
N****g
发帖数: 5172 | 2 ☆─────────────────────────────────────☆
clonee (CLONEE) 于 (Thu Jul 9 20:18:13 2009, 美东) 提到:
发信人: clonee (CLONEE), 信区: shopping
标 题: [求购] officemax 15 off 75 coupon(麻烦联系我Email)
发信站: BBS 未名空间站 (Thu Jul 9 20:17:52 2009, 美东)
麻烦联系我Email：
clonee(at)gmail.com
谢谢！！ |
|
S*****a
发帖数: 63 | 3 【 以下文字转载自 Pharmaceutical 讨论区 】
发信人: Somalia (Anti-Pirates), 信区: Pharmaceutical
标 题: Biologics Discovery and Development Positions Available at Abpro Labs in Woburn, MA
关键字: Biologics,Discovery,Development,Antibody,Computational
发信站: BBS 未名空间站 (Wed May 31 20:28:52 2017, 美东)
Multiple Biologics Discovery and Development Positions Available at Abpro
Labs in Woburn, MA
www.abpro-labs.com
To apply please send resume to [email protected]
1. Scientist/Senior Scientist, Antibody Display Technology
... 阅读全帖 |
|
J*****w
发帖数: 180 | 4 我朋友正在帮国内厂家物色有领导能力的，有创业精神的专家，在国内药厂组建蛋白质
药物开发部。
A leading China Pharma Company has immediate openings for the following four
leadership positions in its Yantai, China and Singapore sites. Please send
your resume to [email protected] if interested.
Title: Head, Upstream Process Development
Functional Description:
Lead, build and develop a high performance Upstream Process Development
group engaged in mammalian cell culture development for commercial
therapeutic proteins, including subclone/clone sel... 阅读全帖 |
|
S*****a
发帖数: 63 | 5 【 以下文字转载自 Pharmaceutical 讨论区 】
发信人: Somalia (Anti-Pirates), 信区: Pharmaceutical
标 题: Biologics Discovery and Development Positions Available at Abpro Labs in Woburn, MA
关键字: Biologics,Discovery,Development,Antibody,Computational
发信站: BBS 未名空间站 (Wed May 31 20:28:52 2017, 美东)
Multiple Biologics Discovery and Development Positions Available at Abpro
Labs in Woburn, MA
www.abpro-labs.com
To apply please send resume to [email protected]
1. Scientist/Senior Scientist, Antibody Display Technology
... 阅读全帖 |
|
S*****a
发帖数: 63 | 6 【 以下文字转载自 Pharmaceutical 讨论区 】
发信人: Somalia (Anti-Pirates), 信区: Pharmaceutical
标 题: Biologics Discovery and Development Positions Available at Abpro Labs in Woburn, MA
关键字: Biologics,Discovery,Development,Antibody,Computational
发信站: BBS 未名空间站 (Wed May 31 20:28:52 2017, 美东)
Multiple Biologics Discovery and Development Positions Available at Abpro
Labs in Woburn, MA
www.abpro-labs.com
To apply please send resume to [email protected]
1. Scientist/Senior Scientist, Antibody Display Technology
... 阅读全帖 |
|
b*s
发帖数: 82482 | 7 晒晒书
很小的时候看过的《Star Wars》，后来陆陆续续看过一些novelization和属于legends
的外传，关于Sith的起源等等，玩过一些星战游戏，但是都没有很系统地钻研这个系列。
最近在跟娃们一起看《Clone Wars》系列，就跟娃一起补了一下课，把《Star Wars》
里面的Canon都又复习了一遍，从图书馆借来了六部蓝光收藏版，把deleted scenes和
制作过程的记录片也看了一遍，包括演员采访，卢卡斯访谈，还有visual artists的访
谈等等。
这卢卡斯不近就靠这个系列吃了一辈子，还养活了一堆人，包括做电影的，写小说的（
以Star Wars fictional universe为题材的小说就不下一百本，当然，大多数都只有
legends的地位，canon还是原来的那几个电影加上《Clone Wars》的电视系列片。
关于星战人物背景和武器装备已经有无数的coffee table books了，最近在图书馆找到
一本星战和地球历史背景的书，跟娃们一起过了一遍。
Star Wars and History
这本书是采访了星战制作人以后，写的关于历... 阅读全帖 |
|
t********7
发帖数: 2691 | 8 I think gay rights ppl will eventually break the legal barrier for human
cloning. I am counting on them:
-------------------------------------------------------
Homosexual couples have the same strong interest as heterosexual couples in
raising children with whom they have biological ties. Genetically related
children permit the continuation of one's lineage and heritage in a way that
is not possible with biologically unrelated children. Moreover, with
genetic parenthood, the same-sex family wou... 阅读全帖 |
|
l**********d
发帖数: 47 | 9 现在继续我昨天的论述。今天我想从个人和国家民族的角度来分析这几件事。我绝对不
敢说我就真的理解了神的心意。说真的，写这个我都是怀着战兢的心，因为神的心意何
其难寻。祂的智慧我只能是象瞎子摸象一样来看，就我所领悟的讲一讲。也许错误难免
，误导人我的罪就大了，求神保守每一个看的人能按正道理解，与你有好的关系。
我说过神的主权是绝对主权，其中包括对道德的期盼。大家知道，不同民族之间，对道
德的定义其实是很不一样的。今天提倡的是普世价值，是在各种价值观的妥协之下形成
的。其实今天的“普世价值”已经受到了基督教价值观的强烈影响。
但神只讲一种价值观，就是神倡导的，其它的都认为是错的。注意不是大家都对，求同
存异。而是只有神对，其他都有偏差。
神的心意是要在人执行这个价值观的，但是世界自从伊甸园事件之后，人自以为拿到了
分别善恶的能力，就开始自己定义“道德”了。分别善恶就是道德。
世界堕落之后，因为自由意志的原因，神不愿强制把意念加给人，而人又不走正路，世
界混乱了，那怎么办呢？
接下来请大家务必带着开放的心来看。注意神看世界与我们看世界的角度是不同的。我
们看世界是以地球的角度，神是以创世主的角... 阅读全帖 |
|
I*******e
发帖数: 1879 | 10 ☆─────────────────────────────────────☆
bruklyn (我爱布鲁克林) 于 (Fri Aug 10 16:35:31 2007) 提到:
protect Object Object.clone()
任何class都subclass Object，就是说任何class都可以access clone()method
但是
public Object myClone(){
Object o1 = new Object();
Object o2 = o1.clone(); //compiler error: clone invisible
o2 = super.clone(); //no error
}
这个super到底是啥type，我的class没有base class，难道super不应该就是Object
type嘛
☆─────────────────────────────────────☆
bruklyn (我爱布鲁克林) 于 (Fri Aug 10 16:38:43 |
|
l*****k
发帖数: 587 | 11 too long to read :)
I think it is possible pet20 have leaky expression, it may kill the cells.
in this case, I think novagen have a cell line that won't have leaky expression.
Instead of cloning disgested fragment, how about do blunt cloning using
novagen's perfect cloning kit? then cut it, at least you won't worry about the
short seq at the restriction site, after cloning,
with multiple clone sites on both side, you
have more choice to put it to pet20, though you have to worry about the problem |
|
g****r
发帖数: 14 | 12 it is possible the gene in the bank has mistakes
but what pcr enzyme did you use, since taq tends to give errors
besides, you may order the clone from IMAG directly, instead of cloning
by yourself, if you are sure the clone is the one that you are going to use
I guess it is cheaper to buy the clone, than do the whole cloning/sequencing by your own |
|
n********k
发帖数: 2818 | 13 I would go for either inducible clones or go viral approach but as transient
approaches...单克隆 is out of dated generally speaking though. and in many
occasions, to have stable clones is not right or simply impossible. and it
depends on reviewers too...I would say most of reviewers will buy viral
approach as transient approach while some hates single clones unless it is
very well characterized with many clones...if one is not careful enough, one
could get single clones of whatever desired phenotyp |
|
n********k
发帖数: 2818 | 14 What an AMAZING woman! seriously!! I vote her to be a next Nobel laureate
and a first Chinese and it is a girl too....
http://english.cas.cn/accessory/twows4th/twowsaward/201006/t20100627_55793.html
Professor Fanyi Zeng was born in 1968. She received her B.A. in University
of California San Diego, CA, USA in 1991. In 2005, she got her Ph.D in
University of Pennsylvania, PA, USA.
Professor Zeng has led a research team that has made substantial and long-
lasting contributions to the developmental ... 阅读全帖 |
|
p*******7
发帖数: 23 | 15 一段时间前，本想‘第一次版上发贴，真心求教’，写了下文。后真的烦不愿再提。今
天读‘中国RA的文，以及回复，希望
分享我的’drama'：
长话短说，RA:发 IF 》12 的一作时我还在本科，不知’科研‘何物。我, 薄厚前活泼
开朗，心直口快（现在，真的知道是
大毛病)，喜欢宏观的可以见效果的。薄厚4年了，文章去工业界多了（朋友言），留学
术界又差着。‘人生的九曲湾
中’放弃追梦. 博后到了距家最近的Lab， 虽然lab focus 与我的Ph.D. training有出入
与此RA曾为朋友，老板3 年前发现课题有潜力， assign 此 RA， 非大陆的。我还开心
，不是因有RA，而是因为是朋
友。（领域真的labor intensive, grad school 与三RA 共事，另一美国博生是我好友
，亦次次出手在大实验：3－5 小
时能拿到要的sample 就很开心乐。这后面才是set-up study.）薄厚Drama开始于这RA
的指定。几乎合作刚开始，朋
友就没了。我傻糊糊的还想继续友谊，修缮关系。直到N 年后被HR告 'forget about
friendship, ... 阅读全帖 |
|
z*t
发帖数: 863 | 16 Beutler得奖，Medzhitov会不会有意见？
http://www.sciencedirect.com/science/article/pii/S1074761309002
Ruslan Medzhitov
This year marks the 20th anniversary of a publication (Janeway, 1989) that
in many ways revolutionized our understanding of the immune system. As part
of the proceedings of the Cold Spring Harbor Symposium on Immune Recognition
, the paper authored by the late Charles A. Janeway, Jr. was not a standard
peer-reviewed publication. In fact, Charlie used to refer to it as “the
best paper I've ... 阅读全帖 |
|
y*********u
发帖数: 183 | 17 请问有哪些commercial available的质粒好用的？
有人用过如下这个质粒么？
Generation of a double-fluorescent double-selectable Cre/loxP indicator
vector for monitoring of intracellular recombination events.
Pfannkuche K, Wunderlich FT, Doss MX, Spitkovsky D, Reppel M, Sachinidis A,
Hescheler J.
Source
Institute for Neurophysiology, Robert Koch Str. 39, 50931 Cologne, Germany.
Abstract
Here we describe the generation of a double-fluorescent Cre/loxP indicator
system. This protocol involves (i) all cloning steps to generate ... 阅读全帖 |
|
S*****a
发帖数: 63 | 18 【 以下文字转载自 Pharmaceutical 讨论区 】
发信人: Somalia (Anti-Pirates), 信区: Pharmaceutical
标 题: Biologics Discovery and Development Positions Available at Abpro Labs in Woburn, MA
关键字: Biologics,Discovery,Development,Antibody,Computational
发信站: BBS 未名空间站 (Wed May 31 20:28:52 2017, 美东)
Multiple Biologics Discovery and Development Positions Available at Abpro
Labs in Woburn, MA
www.abpro-labs.com
To apply please send resume to [email protected]
1. Scientist/Senior Scientist, Antibody Display Technology
... 阅读全帖 |
|
S*****a
发帖数: 63 | 19 Multiple Biologics Discovery and Development Positions Available at Abpro
Labs in Woburn, MA
www.abpro-labs.com
To apply please send resume to [email protected]
1. Scientist/Senior Scientist, Antibody Display Technology
We are seeking an experienced and highly motivated Scientist with strong
hands-on experience in antibody display (phage/yeast) to support discovery
research projects by screening to identify lead candidate therapeutic
antibodies, as well as engineering molecules for improved... 阅读全帖 |
|
发帖数: 1 | 20 克隆的确比复制翻译的好。 但是你说英语没人用 clone a disk 这就彻底错了。英语
可以有 copy a disk. 也可以有 clone a disk. 意义完全不一样。
clone a disk 是指 low level sector-by-sector duplication.
[在 fiveheart (threeheart+2) 的大作中提到：]
:无知岛民觉得什么都是自己的好。同样一个词，大陆的翻译会根据语境变化，完全符
合信达雅的原则，而不是你们一根棒子捅到底：
:太空 空间 大陆两个都用，好坏个屁。space capsule 太空舱，space station 空
间站, 不是空间舱，太空站。一般远的是太空，近的是空间。
:冷气 空调 谁教你的air conditioning只管冷不管热？air只能cold air不能hot
air?
:策略 战略 大陆的战略一词只用于宏观，你背着老婆找小三，那叫策略，不是战略。
:议会 杜马 一提杜马，那只能是俄罗斯。英语也是Duma而不是parliament。而议会
，你
:家也可以开议会。意译永远高于音... 阅读全帖 |
|
m*****k
发帖数: 731 | 21 originally I told her to use hashmap to remember the link between original
node and cloned node,
then she asked for no-extra-space solution, so
I came up with this:
each time after creating the clone node, change the original node's random
pointer to point to the clone. then after the whole linklist is cloned, traverse
again to update the random pointers.
link,她 |
|
p*****2
发帖数: 21240 | 22 最后一题是反着clone BT吧？
将一个bt，mirror一下返回一个新的数，要求不改变原来的树。我跟他说
，我不想做了，头一天坐了6个小时的飞机，他就送我下楼。
class Node:
def __init__(self,val):
self.val=val;
left=None;
right=None;
def clone(root):
if root==None:
return None;
Copy=Node(root.val)
Copy.left=clone(root.right)
Copy.right=clone(root.left)
return Copy |
|
p*****2
发帖数: 21240 | 23 来自主题: JobHunting版 - G 家面经 第一题练了练。没有测试。
class QTreeNode (var color:Int){
val children=new Array[QTreeNode](4)
}
def clone(root:QTreeNode):QTreeNode={
if(root==null) return null
val newNode=new QTreeNode(root.color)
for(i<-0 until 4) newNode.children(i)=clone(root.children(i))
newNode
}
def intersection(left:QTreeNode, right:QTreeNode):QTreeNode={
if(left.color==2 && right.color==2){
val newNode=new QTreeNode(2)
for(i<-0 until 4) newNode.ch... 阅读全帖 |
|
p*****2
发帖数: 21240 | 24 来自主题: JobHunting版 - G 家面经
加了一句，检查这个条件。你觉得够了吗？
def clone(root:QTreeNode):QTreeNode={
if(root==null) return null
val newNode=new QTreeNode(root.color)
for(i<-0 until 4) newNode.children(i)=clone(root.children(i))
newNode
}
def intersection(left:QTreeNode, right:QTreeNode):QTreeNode={
if(left.color==2 && right.color==2){
val newNode=new QTreeNode(2)
for(i<-0 until 4) newNode.children(i)=intersection(left.children
(i), right.children(i))
... 阅读全帖 |
|
p*****2
发帖数: 21240 | 25 来自主题: JobHunting版 - G 家面经 第一题练了练。没有测试。
class QTreeNode (var color:Int){
val children=new Array[QTreeNode](4)
}
def clone(root:QTreeNode):QTreeNode={
if(root==null) return null
val newNode=new QTreeNode(root.color)
for(i<-0 until 4) newNode.children(i)=clone(root.children(i))
newNode
}
def intersection(left:QTreeNode, right:QTreeNode):QTreeNode={
if(left.color==2 && right.color==2){
val newNode=new QTreeNode(2)
for(i<-0 until 4) newNode.ch... 阅读全帖 |
|
p*****2
发帖数: 21240 | 26 来自主题: JobHunting版 - G 家面经
加了一句，检查这个条件。你觉得够了吗？
def clone(root:QTreeNode):QTreeNode={
if(root==null) return null
val newNode=new QTreeNode(root.color)
for(i<-0 until 4) newNode.children(i)=clone(root.children(i))
newNode
}
def intersection(left:QTreeNode, right:QTreeNode):QTreeNode={
if(left.color==2 && right.color==2){
val newNode=new QTreeNode(2)
for(i<-0 until 4) newNode.children(i)=intersection(left.children
(i), right.children(i))
... 阅读全帖 |
|
d*****y
发帖数: 1365 | 27 这道题是大数++的变体
int maxmin(vector digits,int target)
{
if(digits.size()<1) return -1;
int clone = target;
int carry = 0;
int result=0;
sort(digits.begin(),digits.end());
int mul=1;
bool firstdigit= true;
while(clone){
int remainder=clone%10+carry;
vector::iterator iter;
if(firstdigit){... 阅读全帖 |
|
n****3
发帖数: 543 | 28 求转让审稿机会，我的背景如下，附上发表list和email： j*****[email protected]
恳请直接emai直接联系
I have worked as research associate for almost six years in the field of
Molecular and cellular biology and immunology. I have published original
research papers in several peer-reviewed journals such as Blood, Plos one,
Experimental Cell Research, Reproduction, Transgenic Research,
Differentiation, Cell Biology International, and Histology Histopathology.
I have also helped Cellular & Molecular Immunology journal to review two
ma... 阅读全帖 |
|
n****3
发帖数: 543 | 29 求转让审稿机会，我的背景如下，附上发表list和email： j*****[email protected]
恳请直接emai直接联系
I have worked as research associate for almost six years in the field of
Molecular and cellular biology and immunology. I have published original
research papers in several peer-reviewed journals such as Blood, Plos one,
Experimental Cell Research, Reproduction, Transgenic Research,
Differentiation, Cell Biology International, and Histology Histopathology.
I have also helped Cellular & Molecular Immunology journal to review two
ma... 阅读全帖 |
|
n****3
发帖数: 543 | 30 来自主题: Immigration版 - 求审稿机会 求转让审稿机会，我的背景如下，附上发表list和email： j*****[email protected]
恳请直接emai直接联系
I have worked as research associate for almost six years in the field of
Molecular and cellular biology and immunology. I have published original
research papers in several peer-reviewed journals such as Blood, Plos one,
Experimental Cell Research, Reproduction, Transgenic Research,
Differentiation, Cell Biology International, and Histology Histopathology.
I have also helped Cellular & Molecular Immunology journal to review two
ma... 阅读全帖 |
|
n****3
发帖数: 543 | 31 求转让审稿机会，我的背景如下，附上发表list和email： j*****[email protected]
恳请直接emai直接联系
I have worked as research associate for almost six years in the field of
Molecular and cellular biology and immunology. I have published original
research papers in several peer-reviewed journals such as Blood, Plos one,
Experimental Cell Research, Reproduction, Transgenic Research,
Differentiation, Cell Biology International, and Histology Histopathology.
I have also helped Cellular & Molecular Immunology journal to review two
ma... 阅读全帖 |
|
n****3
发帖数: 543 | 32 求转让审稿机会，我的背景如下，附上发表list和email： j*****[email protected]
恳请直接emai直接联系
Jianjun Shi, PhD
Dept. Of Cancer Biology
The Scripps Research Institute
130 Scripps Way, C214
Jupiter, FL 33458, USA
Phone (561)228-3241
Fax 561-228-3058
I have worked as research associate for almost six years in the field of
Molecular and cellular biology and immunology. I have published original
research papers in several peer-reviewed journals such as Blood, Plos one,
Experimental Cell Research, Reproduction, Transgenic Researc... 阅读全帖 |
|
m*******y
发帖数: 14292 | 33 The Infinity AbyssThanos then uses the heroes Thor and Genis-Vell (Captain
Marvel's son) against the death god Walker, who attempts to woo Mistress
Death and then destroy the entity after being rejected.[25] Thanos then
devises a plan to become the All-Father of a new race of Gods created by
himself. Thanos, however, finds himself opposed by the Avengers, former
member Mantis, and her son Quoi, apparently destined to be the Celestial
Messiah. Thanos abandons this plan after having to unite with ... 阅读全帖 |
|
c****e
发帖数: 279 | 34 【 以下文字转载自 SanFrancisco 讨论区 】
发信人: clonee (CLONEE), 信区: SanFrancisco
标 题: 途风网很正规，但怎么会这么便宜，比往返机票都便宜！！ (转载)
发信站: BBS 未名空间站 (Thu Dec 29 17:52:22 2011, 美东)
发信人: clonee (CLONEE), 信区: BackHome
标 题: 途风网很正规，但怎么会这么便宜，比往返机票都便宜！！
发信站: BBS 未名空间站 (Thu Dec 29 17:51:56 2011, 美东)
http://cn.toursforfun.com/14-day-the-ivy-league-famous-universi
有没有诈骗？ |
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b*s
发帖数: 82482 | 35 This was quite a lot of fireworks, but Jobs was not finished. There was
one more big obstacle to tackle: Clones. Jobs felt that Clone Vendors such
as Power Computing were cutting into Apple's high-end market, where they
traditionally made the most profit. Clones had failed to effectively expand
the MacOS market, instead taking customers away from Apple. Jobs remedied
this apparent failure of the Clone experiment by all but pulling its plug.
In early Fall 1997, Apple announced its intention to bu |
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g***l
发帖数: 18555 | 36 COPY过去，DETTACH ATTACH是比较保险的。如果出问题还可以SWITCH BACK。如果能
LIVE接上新SAN的话，调试好CLUSTER，就是DETACH-COPY-ATTACH的时间，不过要注意你
的MASTER, MSDB ,TEMPDB在什么盘上，如果旧SAN还保留的话就是个PRODUCTION DB，应
该很容易。如果旧SAN要关掉，你的MASTER TEMDB MSDB MODEL都要移走，就很麻烦了。
SAN的CLONE也可以，我觉得风险很高，但愿PROGRAM没装在SAN上，要CLONE的话，要随时
准备不WORK，SWITCH BACK，而且在线CLONE的话，会损失数据，离线CLONE，又有DOWNTIME，
其实不如连SERVER也换了得了，另起炉灶，调试好了，搞个RENAME SERVER得了。而且现在
都是64BIT的操作系统，不如升级全做了，WINDOWS 2008+SQL SERVER 2008，一次到位，
用个BACKUP测试好了全换掉，老系统就当DEV好了。单换个STORAGE，风险高，就是硬盘大点，
新SAN可能IO PERFORMANC... 阅读全帖 |
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G**Y
发帖数: 33224 | 37 最好再搞第三块硬盘：
1.把Windows 7 SSD，硬盘3，插到8700上。启动8700自带的Windows 8.1。
2.用clone软件把windows 7 ssd，clone到硬盘3上。
3.确认没问题以后。就可以把Widnows 8.1装到Windows 7 SSD了。
从长远看。最好再买一个硬盘做backup用。硬盘这么便宜。多一两块还是要的的。
如果没有硬盘三：
1.把Windows 7SSD插到8700上。启动8700自带的Windows 8.1。
2.给1TB的硬盘，增加一个分区（windows 8.1， disk management就行）。给Windows
8.1，留200G，搞一个新的700G左右的分区。
3.用clone软件把windows 7 ssd，clone到700G的分区。确认没问题以后。
4. 就可以用Windows 7 SSD重装Widnows 8.1了。 |
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o****i
发帖数: 1706 | 38 public Date clone(){
Date date=null;
try {
date=(Date)super.clone();
} catch (CloneNotSupportedException e) {
// TODO Auto-generated catch block
e.printStackTrace();
}
return date;
}
public Date clone(){
return date.clone();
}
这个是否等效呢？有什么区别吗？谢谢！ |
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t****t
发帖数: 6806 | 39 你如果要改原来的object, 那clone不make sense对吧.
你如果不要改原来的object, 那你clone干嘛?
只有当你确实需要clone的时候,clone才是有用的(比如, 你要以旧object为模板弄个新
的).
我知道你的意思, 你大概说是的STL里那种copy in, copy out的prototype. 不过不知道
问的人是什么意思. |
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r***e
发帖数: 2539 | 40 Thank everyone for the discussion!
As pineseed pointed out, too many topics were discussed here.
But my personal interests are:
1. what is the cell origin of different type of lung cancers?
2. does cell origin or mutation decide tumor types?
3. how to model different type of lung cancer?
HGTPase, it seems you are working in this field (more lung development than
lung cancer?), and I hope you can give your opinions. We can also discuss in
private mails for technique details.
1. ... 阅读全帖 |
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b****r
发帖数: 17995 | 41 问题是肿瘤里一定就是Stem cell (slow cycling)--> transit : amplifying cells (
fast cycling progenitors) 这样一种模式吗。为什么不可以是这么一种模式，类似于
在体外培养耐药菌：
培养皿上种了一个细菌后，它变成一群细菌，每个个体在以同样的速度突变和繁殖，外
来因素如抗叶酸药物磺胺进来后，绝大部分细菌死了，只有偶然突变出耐药性的细菌生
存下来，并且繁殖开来成为relapse的源泉，而且relapse的细胞对原有的化疗相对耐药。
现在所谓CSC而不是single clone expansion的主要证据，如果我理解正确，是数个
clone的expansion而不是细菌常见的单个clone expansion，但是我以为这个非常好理
解，因为化疗药物对肿瘤细胞的杀伤能力基本上是远远逊于抗生素对细菌的杀灭能力的
（因为化疗药物并没有办法真正区分肿瘤细胞和正常繁殖的细胞），所以实际上只是给
肿瘤细胞一个较长周期的外界压力，因而肿瘤细胞有机会在这么长一段时间里以多种不
同的方式进化出抗肿瘤的能力。类似于如果只是以较低浓度... 阅读全帖 |
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H****s
发帖数: 301 | 42 It depends on your purpose and the cell line to be transfected. Generally
speaking, if you cell line is tough enough, puromycin is much easier to work
with. You just need to add a relevant amount of puromycin to the media and
it starts killing.
For neomycin and hygromycin, you have to determine optimal killing
concentration specific to the cell line you will use. To initiate killing,
you have to split cells and supplement with neo/hygro containing media.
Every selective marker has its specific p... 阅读全帖 |
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n******7
发帖数: 12463 | 43
PCR的error有一些，有些clone很奇怪，做了SANGER出来一些不知道哪里的序列。还有
rRNA的序列都弄出来了
我做的assembly，为了保证data size，有些cutoff设定的不是很严格，所以有些区域
的分值不高。还有一些alignment的问题，我发现过几个错误。 我们的目的是发现一些
新的iso，所以ref sequence并不是总有用。
保守没办法，我们的实验pipeline有些assumption，其实是不完全对的。加上我们最后
研究的使protein，这个对mrna序列太敏感了，只好保守一点。除非老板或者合作者愿
意做一些protein level的高通量严重，比如MS之类。。。
我们想发现全长isoform
RNA-seq目前只能研究splicing sites吧？ Cufflinks，scripture什么的出来的
isoform还是根据splicing site预测的，不一定真实存在。 做clone的目的也是为了
克服RNA-seq的这个缺点。我们选了一组gene set做的clone，手上几组数据大概一共测
序了36K的clone |
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b****r
发帖数: 17995 | 44 我是做遗传的对肿瘤涉及不太多
但是平时还是经常关注思考肿瘤，所以也来评一下吧
平心而论，我觉得就这么根据一点缺失啥的找肿瘤病因很容易进死胡同，不是好方向
肿瘤太复杂了，你用什么做材料？如果是原代肿瘤，里面可能是很多细胞clone的混合
物，可能还有大量的stroma cell，你得到的CGH结果到底是什么意思，是有些clone缺
了，有些clone得了copy的综合结果吗？
再如你说的，大部分肿瘤都有genome instability的问题，可以想象，很多gain/loss
其实都是无关紧要的随机事件，从遗传上来说，你发现的gain/loss很有可能只是和真
正的driver连锁了，然后随着clone增生被一起带到后来的细胞群里
另外，做实验难免都要放到体外来，而体外的环境和体内太不一样了，肿瘤在体内要面
对那么多不同的stress，到体外培养条件下都完全不同了，几乎没有stress，唯一的决
定因素就是细胞的增值速度，你养着养着细胞可能就根本不是一群东西了，你再基于这
个做实验，意义何在呢。也许能发点文章，但是对于领域的贡献又何在，这project本
身能走多远，你又学到了一些什么... 阅读全帖 |
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y*****w
发帖数: 146 | 45 sorry, I miscalculated, actually, you should be able to get your
positive
clones if you have around 1-5 in the total cells (means you just need to
sort 5x10^6 cells).
The only potential problem is how you differentiate the positive clones
from
the negative ones. If the sensitivity is good enough, you theoretically
just need one clone. Normally, 5-fold should be safe enough for you to
get
one unless your have a very crappy assay.
It all depends on your assay. I would say 10-fold less of the total... 阅读全帖 |
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s*******e
发帖数: 1389 | 46 有些文章，你觉得data不solid，鄙视它就算了。
可是我当初找博后的时候，读了Ben simons的文章，激动的不行，觉得这才是做表皮干
细胞的好方法，那些wet guy搞什么搞，搞了半天不如人家Simons的一篇文章讲的明白
。于是申请了现在的实验室。
等我来了现在实验室，上手一做才发现，根本不是那么回事啊，后悔来不及了....
Simons的问题是，把生物问题简单化了。比如他在果蝇ISC的文章，Delta staining在
06年刚出来的时候大家都用来作为stem cell的maker，但是后来人们发现Dl在肠道受损
伤的情况下就不特异在stem cell表达了。而他在12年的文章还用Dl作为stem cell的
maker。为什么？就是因为如果你用其他方法一块鉴定stem cell的数目，就不符合他的
Neutral drift的模型了。
还有，就我看过的他的文章，他们做mosaic analysis,不考虑cell migration，也不考
虑clone fusion (这点很复杂，比如他说我10000个cell里面只诱导一个clone， 那么
clone mergin... 阅读全帖 |
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m*********D
发帖数: 1727 | 47 RIP.
钱先生这一走，我最担心的是那个Prasher的饭碗了：
（Zt from WiKi）
Career[edit]
Prasher received his Ph.D. in Biochemistry from the Ohio State University in
1979. From 1979 to 1983, he worked in genetics and biochemistry research at
the University of Georgia, where he identified the gene sequence for
aequorin [1] .[4] He then joined the Biology Department of the Woods Hole
Oceanographic Institution, Woods Hole, Massachusetts where he studied
bioluminescence. In 1988, he received a two-year, $200,000 grant from the... 阅读全帖 |
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a******t
发帖数: 33 | 48 你的主张可以用下列命题概括：
命题1：意识不可以通过clone人复制（A）---> 物质（人脑）不能决定意识(B)
逻辑上，这等价于它的逆否命题：
命题2：物质（人脑）决定意识(非B) ---> 意识可以通过clone人复制 (非A)
对命题2进行讨论可能更容易一些。
从目前的实验科学的理解，clone的人脑是否在物质上absolutely等同于原来的人脑？
我认为没有几个从事clone的人敢这么说。所以命题2不成立。自然其等价命题1也不成
立。 |
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