h***e
发帖数: 146 | 1 http://www.nature.com/nprot/journal/v4/n2/pdf/nprot.2008.227.pdf
Figure 4 | Insertion of a nonselectable DNA fragment by recombineering.
(a) ‘Seamless’ method to insert nonselectable DNA fragment makes use of
selectable markers that can be used for positive as well as negative
selection
(e.g., galK).
Using this method you can change any sequence(From 1 to n bp) in the plasmid
and then homologous recombination into the chromosome. |
|
m******e
发帖数: 18 | 2 MultiSite Gateway® Pro Plus (for flexible cloning of up to four DNA
fragments into a Gateway® Destination vector) from Invitrogen, you can
clone multiple DNA fragments into one vector without using restriction
enzymes or ligases. |
|
e*****n
发帖数: 15 | 3 已经有了whole genome的DNA，要送去测序，测序那边说我要给他们300bp的DNA
fragment, 这样他们才能做library
看了别人，都是sonication来得到DNA fragment的了
可是要怎样的conditions才能得到300bp的呢？
多谢板上大侠指导 |
|
e*****n
发帖数: 15 | 4 我这个应该是DNA methylome sequencing吧
whole genome 的DNA，用invitrogen的那个methyMiner kit enrich methylated DNA，
但是这个kit上说要根据下游的实验用fragment DNA 和bead incubate
测序那边要300bp的DNA fragment，所以我enrich这步就得要300bp的啊 |
|
G********e
发帖数: 528 | 5 I'm not confused either, but find quite a few studies by other people.
Either apoptosis induce H2AX phosphorylation or H2AX phosphorylation
regulate apoptosis.
J Biol Chem. 2000 Mar 31;275(13):9390-5.
Initiation of DNA fragmentation during apoptosis induces phosphorylation of
H2AX histone at serine 139.
DNA Repair (Amst). 2006 May 10;5(5):575-90. Epub 2006 Mar 29.
DNA-PK phosphorylates histone H2AX during apoptotic DNA fragmentation in
mammalian cells.
Molecular Cell, Volume 23, Issue 1, 121-132... 阅读全帖 |
|
w********e
发帖数: 275 | 6 thanks for the information! so do you mean that those fragments are as real
as the fragments following K and R?
Thanks! |
|
g*********d
发帖数: 233 | 7 据英国《每日邮报》1月7日报道，英国科学家找到了“急性癌症”的形成原因：细胞内
的染色体发
生“爆炸”破坏了DNA，从而让人有可能在短时间内患上癌症。相关论文发表于《细胞
》。
传统理论认为癌症是人体经历成千上万次的细胞突变后，慢慢演化的结果。但英国著名
的疾病研究机
构桑格研究所的新发现推翻了这种看法。这暗示了不管人们怎么努力保持身体健康，也
不能保证命运
不会拿他们开玩笑。同时还说明了为什么有些人在体检时根本没发现癌症痕迹，但数月
后突然就被诊
断患上这种疾病了。
桑格学院的科学家是通过研究750个肿瘤的遗传缺陷后得出以上结论的。其中大部分的
案例都与传统
理论相符，染色体的损坏是常年累积的结果。然而，其中至少有1/40的肿瘤不符合“标
准模式”，有的
染色体似乎是在一夜之间遭到破坏的。
参与此项研究的坎贝尔博士称：“测验结果太让我们惊讶了。在一个细胞里面，染色体
经过一次或者是
多次爆炸成为碎片。如果这个细胞开始笨拙地修补，把碎片杂乱的缝合起来，这样就破
坏了原来的
DNA结构，为癌症的快速形成提供了条件。”
坎贝尔博士表示：“这个细胞应该说‘好吧，我放弃’，而不是像对待昂贵的瓷... 阅读全帖 |
|
R****n
发帖数: 708 | 8 I'd like to analyze a DNA fragment from genomic DNA. No PCR, The length is
around 100-300bp. How can I selectively extract the DNA fragment from
genomic DNA.Thanks |
|
n*********m
发帖数: 38 | 9 Look his publication
Conversion of D-ribulose 5-phosphate to D-xylulose 5-phosphate: new insights
from structural and biochemical studies on human RPE.
Liang W, Ouyang S, Shaw N, Joachimiak A, Zhang R, Liu ZJ.
FASEB J. 2011 Feb;25(2):497-504. Epub 2010 Oct 5.
PMID:
20923965
[PubMed - indexed for MEDLINE]
Related citations
2.
Structural basis for the inhibition of human 5,10-methenyltetrahydrofolate
synthetase by N10-substituted folate analogues.
Wu D, Li Y, Song G, Cheng C, Zhang R, Joac... 阅读全帖 |
|
A*******e
发帖数: 284 | 10 呵呵 真是聪明 一下就猜到了，因为手上好几个课题都需要用到这个，所以需要往死里
折腾出来。 载体的问题我解决了，基本没有自连了， 我在载体上插入了一个3k的片段
，然后再用这个改造好的质粒来制备我的载体，这样双切成功能为我所用的载体就只有
6k， 而只有单切可产生自连的有9k，基本容易分开。
现在的问题是vector和fragment的ratio， 我也常用1：2到1：8之间。 但现在我想了
一下，如果载体的长度是片断的200倍， 而它们的摩尔比假定为1：5。 这样在这个连
接体系里面， 一个载体分子 旁边才5个fragment漂浮在左右，一字排开的话， 载体上
每隔1200bp才见到一个片断，这样载体末端和片断相遇的几率是不是非常低了阿； 如
果载体是6k， 片断是1k的话，1：5的比例就能保证相对高的相遇几率了， 我这个推导
对不对？
切开。 |
|
N2
发帖数: 81 | 11 It should work. This kind of method is very efficient. Here are some points
maybe be useful.
1) Gel purify you PCR product.
2)use Nanodrop to estimate the concentration of your PCR products
3) add all fragment 1:1 ratio to vector. Add 100-200ng vector part. It also
depends your fragment size. you may add up to 500ug total DNA per reaction I
guess.
4) you can put 20-40nt overlap between joint points.
How do you screen you positive colonies? Use cloning PCR to check the joints
? you can just scree... 阅读全帖 |
|
z*********8
发帖数: 1203 | 12 我别的都跟你说的那样做的，只有pcr我跑胶看到都是单条带，所以就只有 column
purification。你觉得是我没有割胶纯化的问题么？？
发信人: N2 (Huahua), 信区: Biology
标 题: Re: 怎样用infusion kit克隆多个PCR 片段？？
发信站: BBS 未名空间站 (Tue Jul 26 17:52:12 2011, 美东)
It should work. This kind of method is very efficient. Here are some points
maybe be useful.
1) Gel purify you PCR product.
2)use Nanodrop to estimate the concentration of your PCR products
3) add all fragment 1:1 ratio to vector. Add 100-200ng vector part. It also
depends your fragment size. you may add up to 5... 阅读全帖 |
|
a*******u
发帖数: 19 | 13 N-terminal fragment C-terminal fragment
——————————————— ————————————————
方案1：
引物设计如下
PrimerN_For: A$$$$$$$$$$$$$$
PrimerN_Rev: B**********
PrimerC_For: B&&&&&&&&&&&
PrimerC_Rev: C@@@@@@@@
A,B,C分别表示 不同的内切酶。
用分别将两端PCR下来依次插入到载体中，即N端以空载体为载体插入，然后将C端以N端
插入完毕的质粒做载体插入。这样的问题是多了一个B的内切酶序列，多2个aminos。
deletion mutants允许多2个外源的aminos吗？
方案2：
引物设计如下：
PrimerN_For: A$$$$$$$$$$$$$$
PrimerN_Rev: &&&&**********
PrimerC_For: &&&&&&&&&&&
PrimerC_Rev: C@@... 阅读全帖 |
|
s*********y
发帖数: 387 | 14 co ask.
fragment与B没有binding, 其它3个fragments都与B有binding,这可能吗？ |
|
s**a
发帖数: 293 | 15 完全有可能啊……而且跟片段大小关系不大吧
fragment与B没有binding, 其它3个fragments都与B有binding,这可能吗？ |
|
E********8
发帖数: 22 | 16 possible publication list for people graduated since 2010
118 伊成器
1. N6-methyladenosine in nuclear RNA is a major substrate of the obesity-
associated FTO.
Jia G, Fu Y, Zhao X, Dai Q, Zheng G, Yang Y, Yi C, Lindahl T, Pan T, Yang YG
, He C.
Nat Chem Biol. 2011 Oct 16;7(12):885-7. doi: 10.1038/nchembio.687.
2.Targeting MgrA-mediated virulence regulation in Staphylococcus aureus.
Sun F, Zhou L, Zhao BC, Deng X, Cho H, Yi C, Jian X, Song CX, Luan CH, Bae T
, Li Z, He C.
Chem Biol. 2011 Aug 26;18(8)... 阅读全帖 |
|
|
l**********1
发帖数: 5204 | 18 是吗?
下一步不做啦
ensure efficient recombination in host cell
需要的吧?
please refer:
//www.ncbi.nlm.nih.gov/pmc/articles/PMC155262/
Forty-base-pair overlaps are adequate to ensure efficient recombination in yeast (Hua et al. 1997;
Oldenberg et al. 1997). Hence, we considered it likely that 80-bp recombination linkers, with 40 bp of
vector overlap and 40 bp of fragment overlap, would be adequate to stimulate efficient fragment subcloning |
|
a*******u
发帖数: 19 | 19 Cell的一篇paper
http://www.cell.com/abstract/S0092-8674(09)01506-2
的supplementary
Estimation of absolute number of Jarid2 and Jarid1a molecules per ES cell
nucleus.
Purified recombinant Jarid2 fragment a (Figure 2B, details in Table S9)
containing the epitope
recognized by the Jarid2 antibody was used for protein estimation. The
Jarid1a recombinant
protein containing aa191-290 (cat# H00005927-Q01) and the antibody (cat#
H00005927-A01)
recognizing same fragment were purchased from Abnova. Two-fold s... 阅读全帖 |
|
N2
发帖数: 81 | 20 1)screen 3-5 more colonies and verify the sequence of that site by
sequencing.
2)if there is no correct one, it seems the fragment used for ligation having
that mutation.
3) do a reverse PCR to fix it if you plasmid is not too big.
4)site mutegensis by Quick Change
5)rebuild the plasmid with new fragments from PCR using Phusion DNA
ploymerase
from 3) to 5) find any one which will be easy for you.
good luck! |
|
n********k
发帖数: 2818 | 21 First of all, I could be wrong but I am not sure you are on the right track
trouble-shooting so far...
As some said, this RNA extraction is not that good...That said, for a
cloning work, this may not be a problem at all...different goals require
different quality...As for protein or organic solvent etc, it may not be
that good but it is presumably fine, not horrible I would say..with today's
enzymes(very powerful), it is likely not a problem either...it would be very
bad if you were try to deter... 阅读全帖 |
|
y****6
发帖数: 196 | 22 Thanks for sinister and kaka1000's reply. I labeled the antibody with
fluorescent dyes and quenchers, and I want to digest the antibody to small
fragments to break the FRET between dyes and quenchers. It would be best if
I can digest the antibody into the smallest fragments. Papain may not be the
best choice as it only cut antibody into three pieces. Maybe I can use some
kind of protease mixture? Any suggestion is welcome. |
|
l****y
发帖数: 398 | 23 我觉得趋势就是上面有人说的，同一个run里， 既做MS1，也做MS2；然后用MS2里几个
比较干净的fragment 的 XIC 来定量. 这要求每个peptide被MS2次数足够多，这样XIC
才有足够多的点来define一个峰。
现在机器比较慢，为了让更多不同的peptide被MS2, 人为的用dynamic exclusion减少
同一个peptide被重复MS2的次数；但也造成没法用fragment 的XIC来定量。
所以未来速度比较快的tof仪器应该会重新流行。 |
|
b*******g
发帖数: 1309 | 24 我知道你说的意思
我说的是现在or未来的方法就是把 MRM的transition 直接整合到QTOF里面去，因为
TOFscan的速度越来越快，不再需要MRM那样只scan 一个或两个fragment ion，而是直
接在 MS2里面选择一两个fragment ion。用MS2的 某些 ion的 peak area 来定量，同
时MS2也提供了sequence的信息
另外你说的MS1的信息给我另外一个灵感，哈哈，谢谢。。。。
Ms1 |
|
l**********1
发帖数: 5204 | 25 if to alternative splcing 3'-end multi ployA sites plus that alternative
ORF length are so longer than superscript reverse transcriptase can cover
likes 20kb then only
one choice is use that oligo dt and Multiscibe reverse transcriptase
then PCR different fragment by diffeeernt forward and reverse primer pairs
set then sub-cloning then sequencing then assembly different fragment to
one super long ORF.
Multiscribe |
|
q***7
发帖数: 144 | 26 Use this company's gel extraction kit will allow you get washing buffer-free
linearized vectors/DNA fragments, which will increase your cloning
efficiency.
You can store linearized DNA for over 5 years at -20C and still can ligate
to corresponding DNA fragment for over 90%.
http://www.greenbioresearch.com/gel-extraction-pcr-purification |
|
w********a
发帖数: 324 | 27 如果有好几个insert fragments，你是直接跟vector一起G.A？ 还是先把几个
fragments做G.A.，然后拿这个产物做template得到最终的一个整个的insert，然后再
跟vector 做G.A.？ |
|
f******g
发帖数: 382 | 28 解不出结构的没办法.现在大药厂的基本操作确实是只要有可能,都会去尽力去解.
比如IDH, 发现它有个gain of funtion, 可以作靶点，Agios立刻就找contract
company了结构方面的研究。
象fragment-based drug development,结构就是必须的，而且是不断解药物-蛋白化合
物结构来优化药物。一个例子是abbot的BCL2 inhibitor ABT-XXX 系列。因为是target
蛋白蛋白作用，没筛出什么好药来， 是通过fragment-based drug development找出
来的。
其实大部分的生物研究从长远看还是有用的。只是生物太复杂，产业化在现阶段很难。
学生物做研究确实不挣钱， 下定决心，花点功夫，转行并不难。没有必要把自己辛辛
苦苦做的东西说得那么没用不堪。 |
|
M******g
发帖数: 152 | 29 Just use NEB gibson assembly kit or their newer version NEBuilder hifi DNA
assembly kit. you can clone up to 6 fragments. Make sure use their
competent cells, since the assembly rxn many times is not compatible with
competent cells from other companies such as invitrogen or homemade cells.
you can use NEBuilder software to design your primers: http://nebuilder.neb.com/.
I have assembled 4 fragments and plasmids are normally 7-8 kb. if you have
questions, just call NEB tech support, and they ... 阅读全帖 |
|
g*****x
发帖数: 3283 | 30 你还是没理解bottom-up实验的原理。质谱本身并不明白里边飞的带电粒子是不是
peptide，只是经过fragmentation之后，fragments的pattern和从sequence预测的
pattern很像，所以我们觉得这可能是个peptide。
对于一个已知序列纯化过的蛋白，一般来说可以cover 80-9x％的digest以后的peptide
，但对于比较复杂的体系，比如cell lysate，那么因为peptide太多仪器通量不够，或
者因为非典型salt adduct，数据库序列不全等等原因，很可能会miss掉很多。具体
miss了多少，这个用现有的商业化软件很难evaluate。 |
|
g*****x
发帖数: 3283 | 31 他估计是说用etd，ecd这些fragmentation做top down proteomics。
这个解不了精细结构，但可以在接近native的状态下通过完整蛋白的fragmentation
pattern来了解各个subunit的组装方式。
: 这个是要结合limited proteolysis的吧，属于比较间接的证明。
|
|
s****n
发帖数: 147 | 32 I got some mass spectra, some MS of fragments have 46, 30, 16 difference.
Please some advice, what is fragments are possible difference: 46: COOH?
30: CH3CH2?, 16: O?
Thanks a lot |
|
h*****t
发帖数: 1226 | 33 Tuning File和有没有Fragment没有什么关系．
ETD fragment的效率原本就不如CID, 你要看你的2nd scan event
里面的ETD reaction time, isolation windown width是不是合适．
还有，你要看看你的ETD ??source是不是工作．
MS |
|
b*******g
发帖数: 1309 | 34 对的，这个系统是从LTQ-oribtrap 直接移植过来的，这样做不用重新设计，
只要共用一条生产线，方便简单，成本低。否则不可能只用LTQ-orbi 一半的钱就能买
到Q-Exactive
另外这个CID是High energy CID，or HCD，跟一般的low energy CID 不一样，高能量
对于peptide fragmentation 有帮助
另外，在HCD部分可以很方便的加上ETD collision source （做protein post-
transnational modification analysis），甚至IR，UV等 做不同的fragmentation
研究
这个都极大的丰富仪器在proteomics 跟蛋白结构鉴定方面的应用。这个是一般的
Collision cell 不能做的。 |
|
y*****s
发帖数: 1047 | 35 哪里可以找到Thermo HCD fragmentation的时间和从collision cell转到orbitrap的时
间？还有collision cell的容量。。。orbitrap扫描时间长短本身并不怎么重要，因为
是同时进行不占用时间
我估计activation/fragmentation>10ms,如果加上离子在collision cell， ctrap 和
orbitrap之间传输时间不超过100ms，collision cell的容量足够收集超过0.5s的
parent ion，那效率还是很不错
To
“一台LTQ-orbitrap 100w,一台Q-E才45w，一台普通的QQQ才30w.只做小分子定性跟定量
的话，买Q-E再加QQQ，更便宜，更实用。”
再便宜，我也没钱买，不用帮thermo推销了。。。 |
|
a*****s
发帖数: 838 | 36 I see.. now it seems transformation is the right answer. I read the question
and thought too much :( Thats why I ended up with excluding the most likely
answer first and over analyzed a lot of things.
The transformation happened in the first experiment; the parallel is to tell
you that when the DNA fragments are digested, (therefore no source for tran
sformation), no resistance observed. To answer your question, yes DNase dige
st DNA away. In the parallel experiment, DNA fragments gone, so no tr |
|
c*********5
发帖数: 458 | 37 我妈妈两天前因为摔倒,右肘关节脱臼并骨折.急诊处理后,脱臼复位. 附上复位前后的X
光片,请问,骨折严重吗? 需要进一步治疗吗? 脱臼复位的成功吗? 石膏需要打多久?需
要多久以后进行X光复查? 像这种情况,愈后情况会理想吗? 手臂活动能力会受影响吗?
多谢各位专家的指教!
FINDINGS: 3 views of the right elbow. Dislocation of
the elbow
joint. Several fracture fragments are evident. The donor
sites are not
clearly visible. There is evidence for fracture of at
least to the
olecranon and the radial head.
IMPRESSION:
1. Dislocated... 阅读全帖 |
|
s********o
发帖数: 3319 | 38 restriction enzyme analysis means the 3 viral DNAs are treated with ONE
restriction enzyme and then the digestion products are loaded onto the gel.
a given restriction enzyme only cuts DNA at a specific sequence called
restriction site. usually a DNA fragment has a limited number of restriction
sites for a given enzyme.
in the above case, the linear HSV-1 viral DNA has 10 sites for the enzyme
they used, which produces 11 DNA fragment after the digestion.
D can be a result from a digestion in whi... 阅读全帖 |
|
l**x
发帖数: 110 | 39 知道这个版的专业医生很多，所以占宝地求指点。恳请大家一点给点指导，下一步怎么
办合适？
患者44岁，男，6年前因为肝癌做了肝移植 （符合米兰标准），之后无灾无病好几年。
去年2016年初发现轻微贫血，血色素12，因为当时在服小剂量阿司匹林，于是停服，看
看有没好转。托托拉拉到了年底，还是贫血，血色素10.5。医生说做个内镜。于是在胃
内发现一个肿瘤，内镜医生直觉是GIST。随后CT，但CT却没有异常。4月初第二次胃镜
，GIST依然还在，取样活检表明可能肝癌转移或者是非常罕见的HAC。第二次CT 报告大
小46x35mm，说比较第一次32mm变大了。可能第一次看片医生那天喝多了？肿瘤位置在
Fundus，非常接近GOJ。
AFP好久没有查了，因为肝上的情况一直很好，肝移植前AFP 100多，移植后AFP一直小
于2。上一次至少是2，3年前。也是疏忽了。现在AFP 254. 显然胃上这个要么是转移，
要么是HAC。
下周二医生还要安排一次PETCT，周三见外科医生，估计排除转移外就安排切除了。 求
这里的专家教授指点，这个情况如何是好？下面我贴上最近几个报告。
个人感觉肝移植后5，6年转... 阅读全帖 |
|
c********m
发帖数: 16 | 40 Give you one day to study on the fragmentation of peptide. you will know how
to interpret MS2 of peptides. Nothing difficult. Actually, there are
certain rules applied for all peptide fragmentation. On small molecule, the
spectrum is simpler but the analysis is case by case.
Proteomic is far beyond just protein and peptide characerization. Nowaday it
focuses more on the profiling of a subset of proteonome. Protein
identification is done mostly by search engine instead of by hand. |
|
l****t
发帖数: 25 | 41 alvarado, could you suggest some good articles on class fragmentation, in
particular, the fragmentation of the middle class? Many thanks. |
|
l****t
发帖数: 25 | 42 hi, alvarado, many thanks. Exactly, I am going to explore the consequence
of this class fragmentation on politics. In particular, I am going to
examine the relationship between the class fragmentation, on the one hand,
and political attitudes and participation, on the other hand. |
|
l****t
发帖数: 25 | 43 I have read a couple of books on class fragmentation by sociologists. But,
I have not seen much writing on the effects of class fragmentation on
politics. |
|
i*****s
发帖数: 15215 | 44 艺术家的想象图，索伦龙Sauroniops正在掠食年幼的棘龙，其余的棘龙四散奔逃
Piecing together the past: The fossilised fragment of skull that Andrea Cau
and his colleagues used to identify the newly discovered dinosaur
Jigsaw identification: This image shows how the fragment would have fitted
into the complete skull of the dinosaur. A human skull is shown for
comparison
新浪环球地理讯 北京时间11月8日消息，一种新的掠食性恐龙被命名为Sauroniops
pachytholus，在希腊文中其意思为“索伦之眼”。顾名思义，这个名字来源于《魔戒
》电影中的魔王索伦。索伦龙(暂译)Sauroniops的化石2007年出土于摩洛哥的东南部，
科学家认为它们曾在约9500万年前残酷统治着北非。
研究... 阅读全帖 |
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A****y
发帖数: 337 | 45 一直都念着你昨天的transfer，移植了2颗就有很大的希望！！不要担心fragmentation
，我知道很多姐妹移植了heavyly fragmented的胚胎，结果好孕的！
给你加油！继续祝福你的2ww和Beta test！ |
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w*******y
发帖数: 60932 | 46 Headphone sale at Bestbuy.com stacks with existing 20%-30% off sale.
Sennheiser - RS 180 - $216.99
Sennheiser - RS 170 - $181.99
Sennheiser - RS 160 - $132.99
Monster - PowerBeats - $130.99
Klipsch - Image S4i - $62.99
Klipsch - Image S4 - $50.39
JVC - Black Series - $41.29
Skullcandy - Ink'd Ear Bud - $6.99
JVC - Marshmallow (HAFX34B for Kramer mod?) - $6.99
Hearos - Hearing Protectors (filler, use 2 for 30%) - $2.09
And a bunch more generic earbuds for $7
List of earbud headphones:
http://www.... 阅读全帖 |
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n****o
发帖数: 2276 | 47 来自主题: _WHandFriends版 - 柔肠寸断 今天无聊，拿出工科男生的较真精神来表扬一下信谣传谣。因为这个问题古今中外都有
，因此弄出好多urban/country legends.
话说那天乐子转了个古人装Ｂ的故事，教授马上知道出处。那本叫世说新语的装Ｂ大集就是一典型的把道听途说当经典的例子，但可能正是故事中的不寻常之处，比较能拨
动大家的柔肠，所以成了经典。
举个信谣传谣的例子吧。
东晋的桓温去解放巴蜀，到三峡时，部队里有人捉了个小猿猴，它的妈妈母猿便沿着江
跟着船一路哀号，最后跳上船来，但一上来就死了。后来破开母猿的肚子，发现肠子都
一寸寸地断裂了。桓温听说这件事，就把那个捉小猿的给开除军籍了。 从此就有了柔
肠寸断这个说法。成语词典里给的出处一会说明朝，一会说清朝，是不对的，批评一下。
这里，说的煞有其事，母猿不但一上来就死，而且还有人当法医做尸体解剖，更奇的是
柔肠寸断。学材料的需要仔细研究一下这个现象，搞清楚肠子怎么能变成玻璃。大家都
知道ｗｉｎｄｏｗ系统的defragmentation功能, 这个寸断的Ｅ文估计就是
fragmentation. 我是打死也不会相信有这回事的，但柔肠寸断的说法,fragmentati... 阅读全帖 |
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l*****e
发帖数: 112 | 48 A quote from:
http://www.crossroad.to/articles2/006/migration-2.htm
Dr. Senge also co-authored the report, "Communities of Commitment: The Heart
of Learning Organizations." It highlights the crisis of "fragmentation"
that keeps people from trading "divisive" Biblical views for a more systemic
or holistic perspective.:
"Fragmentation, competition, and reactiveness are not problems to be
solved -- they are frozen patterns of thought to be dissolved. The solvent
we propose is a new way of think |
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s*****t
发帖数: 1994 | 49 Comet Schwassmann-Wachmann 3 Passes the Earth
Credit & Copyright: Thad V'Soske (Cosmotions.com)
Explanation: Rarely does a comet pass this close to Earth. Last week,
dedicated astrofilmographers were able to take advantage of the close approach
of crumbling 73P / Comet Schwassmann-Wachmann 3 to make time-lapse movies of
the fast-moving comet. Large comet fragments passed about 25 times the Moon's
distance from the Earth. The above time lapse movie of Fragment B of Comet
Schwassmann-Wachmann 3 ov |
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b*****t
发帖数: 9671 | 50 下午11:00（31 分钟前）8964 苏联“六四”档案解密：“六四”屠杀三千人从 童言无
忌 戈尔巴乔夫支持六四屠杀。
http://www.google.com/buzz/109245144142018091431/X2G2iCMcpux
Reshared post from Pillar Chang.8964 苏联“六四”档案解密：天安门死3000人 -
诸哥驿站·博客·网摘
作者：封从德 注意这一段对话： Lukyanov reports that the real number of
casualties on Tiananmen Square was 3,000.（Lukyanov：天安门的真实死亡数字是
3000人） Gorbachev: We must be realists. They, like us, have to defend
themselves. Three thousands . . . So what?（戈尔巴乔夫：我们必须现实。他们和
我们一样，必须维护自己。三 千。。。那又怎么样？） 这是“六四”屠杀三千人的最
新证据。此前有三个证据指向三千人这个... 阅读全帖 |
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