s***e
发帖数: 911 | 1 1. 这个重组过程, 一定是在B-form dsDNA和RecA-ssDNA之间完成的吗?
2. RecA也可以直接和dsDNA作用, 形成dsDNA-RecA triplex. 这个结构在重组过程中有
任何作用吗? 比如, 重组可以不可以在dsDNA-RecA 和一个空的ssDNA之间发生?
3. 重组可不可以在dsDNA-RecA和ssDNA-RecA之间发生?
谢谢. |
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L******u
发帖数: 3412 | 2 偶有一个朋友是养work line GSD的~那个娃叫Reca
昨天Reca妈带Reca来我家做客，胖A太么风度了！一副“我的地盘听我的”态度，对人
家管头管脚的。。啧啧，栽面儿啊！Reca童鞋也不甘示弱，进门后就把Ares那一整盆狗
粮都吃完了~~第一次吃的时候阿肥展现绅士风度，让给它吃了~第二次放饭，是给A吃的。
Reca还往上扑着要吃。。直接当面被Ares一口咬在脸上还好它力度控制的好，吓唬吓唬
而已~Reca小妞完全没受伤哈~不过这娃胆子也真大，被咬完还接着往上冲……被它妈拽
回去了。。
是不是很可爱？ |
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L******u
发帖数: 3412 | 3 偶有一个朋友是养work line GSD的~那个娃叫Reca
昨天Reca妈带Reca来我家做客，胖A太么风度了！一副“我的地盘听我的”态度，对人
家管头管脚的。。啧啧，栽面儿啊！Reca童鞋也不甘示弱，进门后就把Ares那一整盆狗
粮都吃完了~~第一次吃的时候阿肥展现绅士风度，让给它吃了~第二次放饭，是给A吃的。
Reca还往上扑着要吃。。直接当面被Ares一口咬在脸上还好它力度控制的好，吓唬吓唬
而已~Reca小妞完全没受伤哈~不过这娃胆子也真大，被咬完还接着往上冲……被它妈拽
回去了。。
是不是很可爱？ |
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e*l
发帖数: 585 | 4 Nature要求注明各个作者的贡献，比如谁conceived the project,谁做的实验，谁写的
文章，
Zhuang文章里的的原话是这么说的：
“M.J.R. and M.B. conceived the STORM imaging concept. M.J.R., DNA imaging
and
data analysis; M.B., RecA imaging and data acquisition. X.Z. supervised the
project.”
idea，做实验和数据分析都是博后的，Zhuang的贡献不明确，如果给她发诺奖确实不能
服人吧，当然牛逼博后带着老板一起拿奖的也是有的，比如李远哲就是靠博后期间的工
作和他的老板一起拿的奖， |
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e*******l
发帖数: 1269 | 5 http://news.consumerreports.org/cars/2011/10/stop-sale-and-reca
The recall affects 159,884 Jetta TDI sedans and SportWagens manufactured
from May 2008 through September 2011 and VW Golf TDIs manufactured from May
2009 through September 2011.
According to online reports, the pulse of the injector operates at the same
frequency as the natural resonance of that injector line, causing it to
become fatigued and fail. The resulting fuel leak can lead to an engine fire
. The injectors in these common-r... 阅读全帖 |
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e*******l
发帖数: 1269 | 6 http://news.consumerreports.org/cars/2011/10/stop-sale-and-reca
The recall affects 159,884 Jetta TDI sedans and SportWagens manufactured
from May 2008 through September 2011 and VW Golf TDIs manufactured from May
2009 through September 2011.
According to online reports, the pulse of the injector operates at the same
frequency as the natural resonance of that injector line, causing it to
become fatigued and fail. The resulting fuel leak can lead to an engine fire
. The injectors in these common-r... 阅读全帖 |
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s********d
发帖数: 503 | 7 朋友不想要了，车况据说应该很好，一点问题也没有，连擦痕也没有，只是VW之前reca
ll过一次，因为我也不懂，所以上来问问。。。看来我ws了 |
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i****a
发帖数: 36252 | 8 来自主题: Automobile版 - 福特逃生门 Ford Escape
http://www.latimes.com/business/money/la-fi-mo-ford-escape-reca
Ford recalls 11,500 new Escapes, tells owners not to drive them
Ford Motor Co. said Thursday it will recall 11,500 of its newly redesigned
and just introduced Escape sport utility vehicles to replace a fuel line,
which could split, leak gasoline and set the vehicle on fire.
The automaker took the unusual precaution of warning owners not to drive the
vehicle and to contact their Ford dealer immediately. |
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x********u
发帖数: 15396 | 18 Diamond Naturals Lamb & Rice 6lb
DLR0101D3XALW Best Before 04 Jan 2013
Diamond Naturals Lamb & Rice 20lb
DLR0101C31XAG Best Before 03 Jan 2013
Diamond Naturals Lamb & Rice 40lb
DLR0101C31XMF Best Before 03 Jan 2013
Diamond Naturals Lamb & Rice 40lb
DLR0101C31XAG Best Before 03 Jan 2013
Diamond Naturals Lamb & Rice 40lb
DLR0101D3... 阅读全帖 |
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c******y
发帖数: 1938 | 19 胖A和reca很有爱哦
我要是冲上去抱了 胖A会不会咬我？
★ 发自iPhone App: ChineseWeb 7.8 |
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L******u
发帖数: 3412 | 20 真的是争宠。。R妈根本没带吃的- -
而且拍这几张时Ares已经在呲牙了，还对Reca GRRR |
|
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L******u
发帖数: 3412 | 22 你看到的是Reca妈身上的袋子，我帮你问问她~ |
|
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L******u
发帖数: 3412 | 24 Reca妈说她在Petco买的，叫treat tote。6.99左右~ |
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c******y
发帖数: 1938 | 25 胖A和reca很有爱哦
我要是冲上去抱了 胖A会不会咬我？
★ 发自iPhone App: ChineseWeb 7.8 |
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L******u
发帖数: 3412 | 26 真的是争宠。。R妈根本没带吃的- -
而且拍这几张时Ares已经在呲牙了，还对Reca GRRR |
|
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L******u
发帖数: 3412 | 28 你看到的是Reca妈身上的袋子，我帮你问问她~ |
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L******u
发帖数: 3412 | 30 Reca妈说她在Petco买的，叫treat tote。6.99左右~ |
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n********k
发帖数: 2818 | 31 have u tried recommendation incompetent cells, never done it myself but I
thought those have been
developed for cloning of this kind...shall be RecA/BCD all minus... |
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H****s
发帖数: 301 | 32 有两个建议：
（1）改双酶切连接为单酶切连接。载体要处理好。
（2）使用DH10B感受态。或者是类似的RecA-菌。
我做的插11kb到5kb的载体上去，基本上都成了。 |
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s********n
发帖数: 2939 | 33 我都是每一次都重新做transformation，平板4度保存，大概可以用2-4周。
大部分Novagen的protein expression strains都是recA+，质粒不稳定，虽然保存在-
80C，接种多次以后难免会有影响。我知道至少有3个negative的例子了。
如果你坚持要这么做，有一个相对low risk的方法，就是用含有葡萄糖的培养基培养，
分装成许多小管，0.5-1 ml （或更大体积），-80C保存，每次拿一管直接培养做表达
，不要多次冻融。
from |
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b**x
发帖数: 8 | 34 Thanks a lot, SmartKevin.
Did you follow the 2007 Nature Biotech paper by Elledge or some other
paper? Did you use RecA- strain or regular strains? If you have a protocol
to spare, 站内通信。
use |
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M***u
发帖数: 165 | 35 像如下标记：
recA1
nprE17
trpC2
等等
前四个字母是基因的名字
请问后边的数字代表什么？怎么感觉是随便编的序列号而已？
比如弄了一堆recA的mutants,给每个编了一堆序列号而已？
谢谢！ |
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n********y
发帖数: 187 | 36 STORM的第一篇文章NATURE METHODS | VOL.3 NO.10 | OCTOBER 2006 Sub-diffraction
-limit
imaging by stochastic optical reconstruction microscopy (STORM)是这样写的
M.J.R. and M.B. conceived the STORM imaging concept. M.J.R., DNA imaging and
data analysis; M.B., RecA imaging and data acquisition. X.Z. supervised the
project. |
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n********k
发帖数: 2818 | 37 make sure it is what you said, if yes, may consider to change bacteria
strains, look for those which can overexpression long repeats DNA, or
inverted repeats, or for retro and lenti viral stuff...essentially those
lost RecABCD, not the only the RecA as the typical strains..if it is true,
this kind of cloning work is not trivial until you figure out right
materials...Or you could beat the nature by brutal force, I heard a guy
picked 20K clones to get one construct/plasmid done, that was before we... 阅读全帖 |
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n********y
发帖数: 187 | 38 好象NATURE系列都有这样的一段8股.
谁能说一下这8股的来龙去脉?
STORM的第一篇文章NATURE METHODS | VOL.3 NO.10 | OCTOBER 2006 Sub-diffraction
-limit imaging by stochastic optical reconstruction microscopy (STORM)是这样
写的
M.J.R. and M.B. conceived the STORM imaging concept. M.J.R., DNA imaging and
data analysis; M.B., RecA imaging and data acquisition. X.Z. supervised the
project.
我看到的是如果老板觉得这IDEA特别亲,就会当自己BABY管屎管尿,最后即使是大老板还
恨不得自己在出身证明上署#1 |
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n********y
发帖数: 187 | 39 庄老板是通讯作者,但是居然IDEA和工作是底下大功的.
STORM的第一篇文章NATURE METHODS | VOL.3 NO.10 | OCTOBER 2006 Sub-diffraction
-limit imaging by stochastic optical reconstruction microscopy (STORM)是这样
写的
M.J.R. and M.B. conceived the STORM imaging concept. M.J.R., DNA imaging and
data analysis; M.B., RecA imaging and data acquisition. X.Z. supervised the
project.
我看到的是如果老板觉得这IDEA特别亲,就会当自己BABY管屎管尿,最后即使是大老板还
恨不得自己在出身证明上署#1 |
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n********y
发帖数: 187 | 40 好象NATURE系列都有这样的一段8股.
谁能说一下这8股的来龙去脉?
STORM的第一篇文章NATURE METHODS | VOL.3 NO.10 | OCTOBER 2006 Sub-diffraction
-limit imaging by stochastic optical reconstruction microscopy (STORM)是这样
写的
M.J.R. and M.B. conceived the STORM imaging concept. M.J.R., DNA imaging and
data analysis; M.B., RecA imaging and data acquisition. X.Z. supervised the
project.
学生肯定是跟庄老板死挣才这样写的,下场如何呢?
我看到的是如果老板觉得这IDEA特别亲,就会当自己BABY管屎管尿,最后即使是大老板还
恨不得自己在出身证明上署#1 |
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n********y
发帖数: 187 | 41 好象NATURE系列都有这样的一段8股.
谁能说一下这8股的来龙去脉?
STORM的第一篇文章NATURE METHODS | VOL.3 NO.10 | OCTOBER 2006 Sub-diffraction
-limit imaging by stochastic optical reconstruction microscopy (STORM)是这样
写的
M.J.R. and M.B. conceived the STORM imaging concept. M.J.R., DNA imaging and
data analysis; M.B., RecA imaging and data acquisition. X.Z. supervised the
project.
学生肯定是跟庄老板死挣才这样写的,下场如何呢?
我看到的是如果老板觉得这IDEA特别亲,就会当自己BABY管屎管尿,最后即使是大老板还
恨不得自己在出身证明上署#1 |
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n********y
发帖数: 187 | 42 如果真是这样,CHU和庄绝对是科学界的匪帮
还到处装清高,把自己装扮成圣人君子---实在下Z
好象NATURE系列都有这样的一段8股.
谁能说一下这8股的来龙去脉?
STORM的第一篇文章NATURE METHODS | VOL.3 NO.10 | OCTOBER 2006 Sub-diffraction
-limit imaging by stochastic optical reconstruction microscopy (STORM)是这样
写的
M.J.R. and M.B. conceived the STORM imaging concept. M.J.R., DNA imaging and
data analysis; M.B., RecA imaging and data acquisition. X.Z. supervised the
project.
学生肯定是跟庄老板死挣才这样写的,下场如何呢?
我看到的是如果老板觉得这IDEA特别亲,就会当自己BABY管屎管尿,最后即使是大老板还
恨不得自己在出身证明上署#1
这STORM的第一篇文章估计是由于从BZ哪儿偷来... 阅读全帖 |
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i****g
发帖数: 3896 | 43 STORM的第一篇文章NATURE METHODS | VOL.3 NO.10 | OCTOBER 2006 Sub-diffraction
-limit imaging by stochastic optical reconstruction microscopy (STORM)是这样
写的
M.J.R. and M.B. conceived the STORM imaging concept. M.J.R., DNA imaging and
data analysis; M.B., RecA imaging and data acquisition. X.Z. supervised the
project.
如果要给这个炸药奖，只怕MJR和MB的贡献比Zhuang还大？
diffraction
and
the |
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g*r
发帖数: 2116 | 44 会不会是这个情况？
Multimers are concatemers/fused products of several plasmids recombined
together. Happens naturally if plasmids are cultivated in recA+ strains of
bacteria. These plasmids are several times as large as the individual
plasmid and therefore run very slowly in agarose (regardless of their
conformation). |
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m******g
发帖数: 467 | 45 基本上就是题目+bulletin point的形式吧，不要纠结我对paper的品味，呵呵。新年快
乐！
PNAS 13.12.03
推荐：
1. Reintroducing domesticated wild mice to sociality induces adaptive
transgenerational effects on MUP expression
promiscuous / monogamous-line
higher MUP (pheromone) expression in pros-line
Male become more attractive
Difference in Mup promoter methylation level
2. High-throughput DNA sequencing errors are reduced by orders of magnitude
using circle sequencing
Previously ~0.1-1x10^-2
Unique library preparation strategy
Copie... 阅读全帖 |
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m******g
发帖数: 467 | 46 基本上就是题目+bulletin point的形式吧，不要纠结我对paper的品味，呵呵。新年快
乐！
PNAS 13.12.03
推荐：
1. Reintroducing domesticated wild mice to sociality induces adaptive
transgenerational effects on MUP expression
promiscuous / monogamous-line
higher MUP (pheromone) expression in pros-line
Male become more attractive
Difference in Mup promoter methylation level
2. High-throughput DNA sequencing errors are reduced by orders of magnitude
using circle sequencing
Previously ~0.1-1x10^-2
Unique library preparation strategy
Copie... 阅读全帖 |
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j*b
发帖数: 341 | 47 Sub-diffraction-limit imaging by stochastic optical reconstruction
microscopy (STORM)
Michael J Rust1, 5, Mark Bates2, 5 & Xiaowei Zhuang1, 3, 4
Author Contributions
M.J.R. and M.B. conceived the STORM imaging concept. M.J.R., DNA imaging and
data analysis; M.B., RecA imaging and data acquisition. X.Z. supervised the
project.
这才是真正原因，其他都是扯淡！庄太 nice 了。 |
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s***e
发帖数: 911 | 48 你说的这种是特定的重要生物过程信号通路里的Mechanoisensing. YAP/TAZ的
subcellular localization肯定受力调控，这个是已经实验确定的。Notch pathway
notch receptor cleavage也已经知道是受力调控的。刚才我顺手google, 发现也有很
多文献报道wnt pathway也直接受力调控。这些实验表明力可以是调控重要生物过程的
重要因素，不代表是必要因素。所以你的结果其实和Hippo mechanosensing没有显著茅
盾。对了，能分享一下你怎么通过knockout模型来讨论hippo和mechanosensing的联系
的？还想请教一下YAP在细胞质内是和那些蛋白相互作用？我对hippo mechanosensing
很感兴趣，但是对hippo相关的生物基本不了解。正在自学。
除了这些有名字的信号通路，mechanosensing在cell-ECM连接，cell-cell连接, cell
migration, tissue maintenance, 离子通道, actin stabilization, mus... 阅读全帖 |
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j****n
发帖数: 3370 | 49 隔壁实验室也有人这么操作 以前一直觉得大部分cloning host都是recA敲除，重组的
效率会很低
这个最多可以搞几个片段？我们实验室的一般都会有3个及以上片段
另外就是PCR，如果不是单一条带，克隆的准确性有多高？尤其是有好几个片段的情况下 |
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b**s
发帖数: 589 | 50 The bottom-up approach to fabricating nanometer-size electronic devices
requires molecular building blocks to recognize each other correctly and
self-assemble. Biomolecules are attractive candidates, offering binding
specificity and accurate enzymatic processing.
Scientists at the Technion-Israel and Institute of Technology, Haifa, used
these properties of DNA and RecA protein in a process they call molecular
lithography. They have created conductive wires with insulating gaps by
coating p |
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