S*******i
发帖数: 2018 | 1 现在主媒上思维还算正常的文章已经不多见了：
http://www.wsj.com/articles/the-manafort-indictment-1509402445
Special Counsel Robert Mueller indicted former Trump campaign chairman Paul
Manafort for tax fraud on Monday, and the main charge against Donald Trump
is poor judgment for hiring the notorious Beltway operator.
The indictment accuses Mr. Manafort (and business partner Richard Gates ) of
funneling money from a pro-Russia party in Ukraine into offshore shell
companies and bank accounts. They then allegedly used th... 阅读全帖 |
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s********t
发帖数: 930 | 2 CNN: Trump 'seething' as Mueller probe reaches former aides
Washington (CNN)The dramatic intensification of the Russia investigation on
Monday reignited President Donald Trump's fury at the controversy clouding
his presidency, prompting his aides to urgently advise him against lashing
out at special counsel Robert Mueller as they work to revive a halting
policy agenda.
Watching the developments unfold on the large television screens installed
in his private residence, Trump was "seething," accor... 阅读全帖 |
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j*********r
发帖数: 24733 | 3 Harvard faces DOJ probe over its affirmative-action policies
The U.S. Department of Justice has opened an investigation into the use of
race in Harvard University’s admissions practices and has accused the
university of failing to cooperate with the probe, according to documents
reviewed by The Wall Street Journal.
The Justice Department is investigating complaints that formed the basis of
a federal civil lawsuit filed in 2014 in Boston, according to the documents.
That suit alleges Harvard inte... 阅读全帖 |
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j*********r
发帖数: 24733 | 4 G-man told FBI mistress there was nothing to Trump-Russia collusion – but
joined probe anyway
New text messages allegedly reveal that controversial FBI official Peter
Strzok was hesitant about joining Special Counsel Robert Mueller’s Russia
investigation team because of his “gut sense” there would be “no big
there there.”
Strzok and FBI colleague Lisa Page, who were romantically involved, are
under fire for trading anti-Trump messages during the 2016 race.
But Senate Homeland Security Committee ... 阅读全帖 |
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s********e
发帖数: 1503 | 5 看门狗把头像都换了，sad。
：G-man told FBI mistress there was nothing to Trump-Russia collusion – but
：joined probe anyway
：New text messages allegedly reveal that controversial FBI official Peter
：Strzok was hesitant about joining Special Counsel Robert Mueller’s Russia
：investigation team because of his “gut sense” there would be “no big
：there there.”
：Strzok and FBI colleague Lisa Page, who were romantically involved, are
：under fire for trading anti-Trump messages during the 2016 race.
：But Senate Hom... 阅读全帖 |
|
发帖数: 1 | 6 艷星丹尼爾斯（左）在ABC節目談她與川普緋聞及封口費。（美聯社）
川普緋聞女主角、色情片女星丹尼爾斯（Stormy Daniels）17日公布2011年曾在拉斯維
加斯停車場對她出言恐嚇的男子素描畫像，希望社會大眾協助指認男子身份。丹尼爾斯
的律師艾凡亞迪（Michael Avenatti）表示，提供線索讓這名男子身份得以釐清的民眾
，將可獲得10萬美元報酬。
丹尼爾斯指稱，2006年時曾與尚未踏入政壇且與第三任妻子梅蘭妮亞新婚不久的紐約房
地產大亨川普有過婚外情。她也表示，2016年總統大選之前，川普私人律師柯恩（
Michael Cohen）給了她13萬美元「封口費」，要求她不得洩露與川普的桃色關係。到
目前為止，川普總統對於丹尼爾斯的說詞均予否認。
丹尼爾斯17日上午參加美國廣播公司（ABC）晨間談話節目「觀點」（The View）時，
公布了2011年曾在拉斯維加斯一處停車場恐嚇她的男子素描畫像。她說，這名男子要她
不要對外透露與川普之間的關係。
她表示，當年遭到這名男子威脅之後，沒有立即報警的原因是擔心如果報了警，她與川
普的關係就會因此曝光，而且她也處於恐懼之中。
丹尼爾斯... 阅读全帖 |
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l*h
发帖数: 4124 | 7 Hillary Clinton的前首席策士Mark Penn的看法跟我们的一样，deep state和HRC的竞
选团队构陷Donald Trump：
Stopping Robert Mueller to protect us all
Mark Penn
The “deep state” is in a deep state of desperation. With little time left
before the Justice Department inspector general’s report becomes public,
and with special counsel Robert Mueller having failed to bring down Donald
Trump after a year of trying, they know a reckoning is coming.
At this point, there is little doubt that the highest echelons of the FBI
and the Justi... 阅读全帖 |
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b*****d
发帖数: 61690 | 8 The lawyer for Patrick J. Smyth, a former classmate of Supreme Court nominee
Brett Kavanaugh, confirmed on Monday that his client has spoken to the FBI
as part of its probe into allegations of sexual misconduct against Kavanaugh.
Eric Bruce, the attorney representing Smyth, said in a statement that Smyth
reiterated his previous claim that he does not remember the party where
Christine Blasey Ford claims Kavanaugh attacked her.
ord has named Smyth as another attendee at the 1982 party.
"Patrick J... 阅读全帖 |
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c*******e
发帖数: 5818 | 9 感觉 FBI 几乎就是半个 H /民主党的竞选助手，这样还是输了！
老话讲：老天有眼！！
Lawyer for Clinton campaign and DNC gave FBI documents for Russia probe,
sources say
A top lawyer working with the Democratic National Committee and Clinton
campaign contacted the FBI’s general counsel in late 2016 and provided
documents for the Russia probe as federal investigators prepared a
surveillance warrant for Trump campaign aide Carter Page, sources close to a
congressional investigation told Fox News, citing new testimony.
https://www.foxnew... 阅读全帖 |
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f*******e
发帖数: 3433 | 10 The Senate Judiciary Committee released an executive summary of the FBI's
supplemental background report into Supreme Court nominee Brett Kavanaugh
late Thursday, which key swing-vote senators vowed they would continue to
review Friday ahead of a major vote on his confirmation.
According to the summary, released by the Republican committee majority, FBI
agents interviewed 10 people and reached out to 11, focusing only on
witnesses with potential first-hand knowledge of alleged sexual misconduct
... 阅读全帖 |
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d*********r
发帖数: 11979 | 11 Add a comment (474 comments )
Email | Start Following | Print | Facebook Share on Facebook |
View full size
Investigators work inside a Ruby Hill home allegedly under federal
investigation. new
Investigators work inside a Ruby Hill home allegedly under federal
investigation.Credit SusanC.Schena
Photos (6)
Home on Germano Way in Ruby Hill is under Homeland Security investigation.
Credit SusanC.Schena http://o1.aolcdn.com/dims-shared/dims3/PATCH/resize/273x203/http://hss-prod.hss.aol.com/hss/s... 阅读全帖 |
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p*l
发帖数: 1359 | 12 一个合作项目，对方是个外校的教授，资历比较老。昨天他一封email，无端指责我们
，搞得我很不高兴。
对方做一个新的生化荧光probe，想用在细胞的显微成像上。一个学生做了六年了，卡
在了成像验证probe的有效性上。他们的probe扔到细胞里以后，现象太复杂，传统的荧
光显微镜看到的是一团浆糊。我是做新荧光成像方法的。他们通过合作的关系，知道了
我，用我的新成像方法一试，现象能说清楚了。他们就带着他们的细胞飞到我这里做了
两次，共二十天的实验。
当初开始的时候，我就说，我们的新成像方法还没发文章，所以我们得拿他们的一点数
据，先发个文章，把成像方法建立起来，这样他们的生化文章才发得出去。对方老板也
同意了，我也就没和他要他们文章的并列第一作者，想着反正我们有自己的文章，第二
片生化的文章，第二作者也就罢了。昨天我把稿子发给他，他居然变卦了，很不高兴得
说他们在写大文章，我们的文章得等他们的大文章发出去才能发。问题是，他们的数据
用了一个崭新的成像方法，如果到时候reviewer问起来，他们肯定会要我把技术设计都
填进去，那我自己的文章，就发不出去了。这个仪器，我也做了6年，我不愿意这... 阅读全帖 |
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H****y
发帖数: 2992 | 13 Interview with Dr. Xi Yin
--- By Fiona Rawsontile, Sept 2015
This interview was inspired by an earlier interview of Dr. Yin I saw on the
Internet, which made me think that we can’t expect someone who normally
writes for entertainment to understand a physicist. To “provoke” a
scientist, we need another scientist. So I volunteered (to myself) and sent
an invitation to Dr. Yin, who was recently promoted to Professor in Physics
at Harvard Uni... 阅读全帖 |
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S*******t
发帖数: 3956 | 14 （作为年仅31岁就晋升哈佛正教授的青年才俊，尹希教授最近得到了很多关注。其中一
篇关于他的专访进入了一名同是美国大学华裔教授的女科学家的视线。因为觉得这样的
采访不过瘾，她突发奇想，想亲自采访一下尹希。于是就有了这个我们今天看到的两名
科学家之间的对话。这位女教授使用了化名，这是她出版自己几部科幻小说的笔名。对
，没错，这位女教授同时也是一个女文青，已经写了几本书，还都是英文，Amazon 就
能买到。感兴趣的不妨找来看看。很可能有一天，她也会成为传奇式的人物。那今天这
篇采访就会成为另一段佳话。）
Interview with Dr. Xi Yin
--- By Fiona Rawsontile, Sept 2015
Translated by Slow Rabbit
This interview was inspired by an earlier interview of Dr. Yin I saw on the
Internet, which made me think that we can’t expect some... 阅读全帖 |
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g***r
发帖数: 285 | 15
interesting, when the term of ablation mentioned, either radiofrequency
abalation, to me, it means to apply a probe that emmits heat to burn the
tissue, or cryoabaltion, the probe that emeits cold or whatever to freeze
the tissue; either case you have to place the probe directly into the tumor,
which has to be invasive, though less so comparing a lobectomy,
even in chemo-embolization, like they do for liver mets, you still have to
insert the catheter through the tumor site to deliver chemo agen |
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F**D
发帖数: 6472 | 16 Contact Info:
Thomas E. Beechem III
Senior Member of the Technical Staff
Nanomaterials Sciences Department
Email: t*****[email protected]
Phone: (505) 284-3503
Position Opening:
Postdoctoral Appointee – Raman Spectroscopy for Thermal Property Characteri
zation
Description:
The thermal response of a system now increasingly defines the total performa
nce of applications on the whole. As a consequence, the material properties
defining this thermal response must be accurately known and quantified in so
m... 阅读全帖 |
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k********n
发帖数: 18523 | 17 苹果公司怎么可以让这样的事情发生？
NEW YORK (AP) -- Federal prosecutors in Manhattan broadened their insider
trading crackdown Thursday, arresting four people on charges alleging that
so-called "expert consultants" revealed secrets about Apple Inc.'s iPhone
and other technology products to hedge funds seeking a trading edge on
quarterly earnings reports.
The latest probe targeted Primary Global Research, a Mountain View, Calif.-
based firm that offered consulting services to investors on industry trends,
issues ... 阅读全帖 |
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F****s
发帖数: 3761 | 18 Tesla Is Facing U.S. Criminal Probe Over Elon Musk Statements
By Tom Schoenberg and Matt Robinson
September 18, 2018
Tesla Inc. is under investigation by the Justice Department over public
statements made by the company and Chief Executive Officer Elon Musk,
according to two people familiar with the matter. The criminal probe is
running alongside a previously reported civil inquiry by securities
regulators.
Federal prosecutors opened a fraud investigation after Musk tweeted last
month that he w... 阅读全帖 |
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i*****s
发帖数: 15215 | 19 Man deported from Canada admits crimes: China
Reuters – Fri, Dec 30, 2011.
BEIJING (Reuters) - China's most wanted man, who was deported from Canada in
July after a decades-long legal battle, has admitted to his crimes and will
now be handed over to prosecutors, state media reported on Friday.
Beijing had sought the deportation of Lai Changxing for years, accusing him
of running a multi-billion dollar smuggling ring in the southeastern city of
Xiamen in the 1990s in one of China's biggest politi... 阅读全帖 |
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H*****r
发帖数: 764 | 20 http://sports.yahoo.com/investigations/news?slug=ys-miami_viola
CORAL GABLES, Fla. – The NCAA informed University of Miami administrators
it will consider invoking its “willful violators” clause and make an
exception to the traditional four-year statute of limitations in the Nevin
Shapiro case, a university source told Yahoo! Sports.
Traditionally, the NCAA’s bylaws would only allow it to sanction the
Hurricanes for infractions that occurred during the four years prior to
receiving a letter of i... 阅读全帖 |
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g***y
发帖数: 12 | 21 a/ 裸bcrush （针对t，偶尔针对z，p）
8bp9bf
8probe，9probe同时探路。8probe在人家家里逛一圈，然后在
人家眼皮底下溜走。9probe偷偷溜进别人家里角落。
13bp 看情况，15家里1bc，敌人家里2bc。（优先bc，可以停probe）。
2bc好（一定要等好），向前造bc，需要可以打到敌人气矿。
然后就是家里补probe，bc，敌人家里bc，bp的时间。
——————————————————————
b、偷bc战术 （针对t，偶尔针对z，p）
8bp10bg 在自家角落里10bp12bf。
8号农民去敌人家探路，然后离开，再偷偷折回角落。
再敌人家13bp，16probe的时候2bc（停probe），看情况，家里再补bc。
——————————————————————
c/ 单bg，bc封口 (针对t）
8bp9bg10bf，在有虫族协助下，直接探路农民bp封口。后续发展
有2bc～5bc封口（进去侦察t族发展方向），如果t族3兵营以上mm，需要补到5bc
如果2兵营mm，tank补到3bc。如果速tank晚ba，就2bc足够。
自家策略，速气矿 |
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h*****0
发帖数: 4889 | 22 弹射总是优先向左（在弹射范围内的）。这点是没错的。
但是问题在于pylon虽然个头大，但周围是有大量空隙的。当一个单位处于射程范围内
最左边界处，则此单位会100%的被弹射。而pylon不管怎么摆，这个点（大约离被攻击
BC中心往左3个建筑格，即1.5个pylon身位）总是空的，有可能被单位占据。
如果想要实现“不论单位怎么跑，当飞龙攻击BC时总不会溅射到”的美梦，唯一可能恐
怕是用probe去占据那个点并hold住。不过只有一个probe，要造probe还要先造nexus，
非常的杯剧。此方案不太现实。
游戏中相邻玩家间隔大约为3个半pylon，即7个建筑格，如果假定舒服安全的操作需要
至少右边3个建筑格的话，那右侧玩家“不能在原来BC左侧更左的位置造建筑”。可以
用前面攻略里的偏左竖直向下两个pylon的方式，注意自己的兵不要到这两个pylon左侧
去就行。
图示为：（图中4个字母为一个建筑）
CC
CC
PP
PP
PP
PP
要注意很多人用的如下图方式建筑，是有潜在弹射到左侧玩家的风险的：
CC
CC
PP
PP
PP
PP |
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d**********t
发帖数: 20415 | 23 不好用吧,scv打水晶打半天，probe轻松在加一个，你就白忙活了
probe被你打到，跑几圈护顿又好了...而且P一个probe放好多建筑的机制，靠scv很难
打下来
我觉得先补地堡，如果射程差点，就农民修，憋出一个mara就没事了 |
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k****w
发帖数: 3753 | 24 General Micro Tips:
1. Are your zealots in front of your stalkers?
2. Are your low-health units in the back of your army before you attack?
3. Are your immortals attacking armored units?
4. Are your colossus attacking clumped up units? If the entire enemy army is
clumped, than focus on attacking light units (marines, hydras, zealots)
5. When defending your ramp,
A. Are you units in a concave at the top of your ramp?
B. Are you cutting his army into bite-size pieces?
C. Are your... 阅读全帖 |
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h*****0
发帖数: 4889 | 25 xx 没死之前，probe移动围枪兵，枪兵没有太好的办法。除非不进P家。
你单气，没开2矿，也没停农民，1XX 1追猎就all in过来了？那肯定就是拉所有农民拼
了。2BG够了。
如果按我的开局，先不造BG，等钱开基地，这过程中在侦察，如果近点或者对点直接就
看到了，除非较大的4人图最后一点，那我会先下基地，然后几乎同时BG。此时有点小
危险，但马上取消基地补另一个BG，刷XX，同时（钱很足，因为BG晚）开气，农民不停
，T来的时候会有两XX，另两个在造，BY也差不多好了，好了有气出哨兵或者追猎，多
余的水晶全刷成XX，打的时候拖出所有probe，一定要断掉T的后路围上打，就没问题。
scv换光XX，新的XX出来了。只要还有XX在，probe打枪兵很猛的。
你是正常科技开（但又有想开矿的意思，可以时间上稍慢一点），没有先造XX，所以兵
力反而不如我这种。当然你追猎早，操作好可以用追猎搞定，那优势就天大了。
我觉得T这样拉过来的最多也就14个scv，P家里20个农民，肯定能拼。 |
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d**********t
发帖数: 20415 | 26 本来想圣诞写的，实在是懒，拖到现在...
还是那句话，本人菜鸟，理解不足之处，欢迎高玩来拍~
P的战术整体给我感觉是猜对了，很凶猛，猜错了，很悲剧，因为兵种特点都太鲜明，
针对上了可以暴力的一波A死人，但是为啥说猜呢，因为P的侦查能力么，大家都懂的..
.T有雷达,Z有狗占塔，P只好等OB出来，前期的侦查基本是很难的。我用P打天梯基本用
大招，因为玩运营一路跌到黄金，果断不能忍，上套餐，基本白金之前胜率能到80%+，
即使到了白金之后，也能保持70%以上，钻石之后套餐威力就一般了，不过PvZ的那招还
是很好用（下面具体讲）
先说说PvT吧，这个不好打，因为T能13甩的原因，对P的哨兵力场要求相当高，像我这
种框一下A的选手来说比较痛苦，那就索性单矿一波吧...PvT的单矿一波，我主要是5BG
，3BGVS，3BGVR巨像，4BGVR运输机，速闪烁追猎，其他像速DT和3BGVR带不朽也不错，
但是我用的比较少，DT主要是怕女妖真善美之类的，这年头在雷达面前，DT要破口也不
容易，基本只能用来拖延对手2矿打运营，3BGVR带不朽主要是我不朽操作太烂，经常卡
来卡去。选择什么一波战术，基本取... 阅读全帖 |
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b******s
发帖数: 695 | 27 http://slickdeals.net/f/3749468-Free-WIFI-on-Delta-Airline-Enti
DO NOT DO THIS!!! I did this on my last flight and they caught me 2 hours
into the flight. They said it was fraud and had the US marshal place me in
hand cuffs. I had to sit in the cargo hold for 3 hours being interrogated.
They wanted to know where I had learned how to do this. They beat me. They
stripped me. They probed me. When we landed they put a sack over my head a
drove me to some detention center. I spend 3 months there with... 阅读全帖 |
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c****t
发帖数: 19049 | 28 AN ORISON OF SONMI~451
On behalf of my ministry, thank you for agreeing to this final interview.
Please remember, this
isn’t an interrogation, or a trial. Your version of the truth is the only
one that matters.
TRUTH IS SINGULAR. ITS “VERSIONS” ARE MISTRUTHS.
… Good. Ordinarily, I begin by asking prisoners to recall their earliest
memories to provide a
context for corpocratic historians of the future. Fabricants have no
earliest memories,
Archivist. One twenty-four-hour cycle in Papa Song’s is i... 阅读全帖 |
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S*******t
发帖数: 3956 | 29 【 以下文字转载自 Faculty 讨论区 】
发信人: SlowRabit (慢吞吞的小白兔), 信区: Faculty
标 题: 翻译：高妹对尹希教授的专访
发信站: BBS 未名空间站 (Sat Sep 26 11:16:17 2015, 美东)
（作为年仅31岁就晋升哈佛正教授的青年才俊，尹希教授最近得到了很多关注。其中一
篇关于他的专访进入了一名同是美国大学华裔教授的女科学家的视线。因为觉得这样的
采访不过瘾，她突发奇想，想亲自采访一下尹希。于是就有了这个我们今天看到的两名
科学家之间的对话。这位女教授使用了化名，这是她出版自己几部科幻小说的笔名。对
，没错，这位女教授同时也是一个女文青，已经写了几本书，还都是英文，Amazon 就
能买到。感兴趣的不妨找来看看。很可能有一天，她也会成为传奇式的人物。那今天这
篇采访就会成为另一段佳话。）
Interview with Dr. Xi Yin
--- By Fiona Rawsontile, Sept 2015
Translated by Slow Rabbit
Thi... 阅读全帖 |
|
|
x****u
发帖数: 12955 | 31
The key to that incident is that the probes broke the skin and reached blood
stream, and that the probes were on each side of the body, requiring the
current to pass through the heart.
With a cellphone charger, there are no sharp probes to break your skin, and
even if you somehow managed that, your heart is not in the path of the
current. |
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e***y
发帖数: 1152 | 32 好像算了个吧星期了，才算到：
...
Probing: '0iTiYX' [1776 pwds/sec]
Probing: '0iTknb' [1782 pwds/sec]
Probing: '0iTlLm' [1781 pwds/sec]
...
估计把六位数算完得等到圣诞节啦。。。 |
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S*A
发帖数: 7142 | 33
不是很理解，你是要自己设计个板子还是用现成比较接近的就可以了？
你说得这个 pogoplug V4 就有，amazon 上面 $7。
100/
Kernel 启动到 User space 我猜customize kernel 2 秒钟应该可以。
从 USB storage 上读就慢了。还有启动网络也慢，要 link negotiate.
DHCP 等等。但是你这种设备如果就是 Kernel 启动了，网络不能用
大概也不行。
http( 不
我觉得，你现在完全没有概念时间到底慢在什么地方。
花点时间看看 dmesg 知道各个模块各自花多少时间。
probe 一般不是多线程的，进入 user space modprobe 可以多线程。
systemd 就是干这个。 UART 通信非常简单，不需要 probe。 那个几乎
没有时间花在 probe 上面。数据发出去要点时间。
如果我是你可能就先拿那个 pogoplug V4 mobile 试试，有 128M RAM，
128M flash。带 UART， Gigabit ethernet, USB controller, SD card.
... 阅读全帖 |
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S*A
发帖数: 7142 | 34
不是很理解，你是要自己设计个板子还是用现成比较接近的就可以了？
你说得这个 pogoplug V4 就有，amazon 上面 $7。
100/
Kernel 启动到 User space 我猜customize kernel 2 秒钟应该可以。
从 USB storage 上读就慢了。还有启动网络也慢，要 link negotiate.
DHCP 等等。但是你这种设备如果就是 Kernel 启动了，网络不能用
大概也不行。
http( 不
我觉得，你现在完全没有概念时间到底慢在什么地方。
花点时间看看 dmesg 知道各个模块各自花多少时间。
probe 一般不是多线程的，进入 user space modprobe 可以多线程。
systemd 就是干这个。 UART 通信非常简单，不需要 probe。 那个几乎
没有时间花在 probe 上面。数据发出去要点时间。
如果我是你可能就先拿那个 pogoplug V4 mobile 试试，有 128M RAM，
128M flash。带 UART， Gigabit ethernet, USB controller, SD card.
... 阅读全帖 |
|
b***i
发帖数: 3043 | 35 新的信息
大神帮看看那里可以提高？
[ 0.128265] omap_i2c omap_i2c.1: bus 1 rev2.4.0 at 100 kHz
[ 0.129791] tps65910 1-002d: JTAGREVNUM 0x1
[ 0.133728] print_constraints: VRTC:
[ 0.135223] print_constraints: VIO: at 1500 mV
[ 0.137603] print_constraints: VDD1: 600 <--> 1500 mV at 1100 mV normal
[ 0.139923] print_constraints: VDD2: at 1100 mV
[ 0.140960] print_constraints: VDD3: 5000 mV
[ 0.142425] print_constraints: VDIG1: at 1800 mV
[ 0.143890] print_constraints: VDIG2: at 1800 mV
[ ... 阅读全帖 |
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c*****t
发帖数: 1879 | 36 1. before installing a new scsi device, probe scsi to locate free
scsi id, then shut down computer
2. set the harddisk the target # (scsi id). attach it to computer
3. start system and probe again.
I think that probe-scsi-all is the command. |
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M****e
发帖数: 70 | 37 my point is to make sure that you have transfer of good
quality and quantity of RNA onto the blotting material.
the washing stringency for your experiment is not quite
high (i.e., 42C). i think the problem is mainly due to
bad RNA or probe. if there is a lot of radioactive
nucleotides incorporated into the probe, you should
detect very hot signal after purification. denature the
probe before use. GAPDH is a housekeeping gene that is
typically used for normalization of load. i suggest you
read a |
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s*******e
发帖数: 740 | 38 哦，忘了提醒你一下
关于第三点，一般人们并不用target序列做probe，而要把probe选在target区域外
这是因为你的序列也可以randomly整合到genomic DNA中去，而你只想要homologous
recombinate的产物，所以不要用target序列做probe，否则有很多假的positve
Good luck! |
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v***a
发帖数: 1242 | 39 测TER (transepithelial resistance)的时候，大家有遇到过读数老是漂移不定的情况
吗？
忽高忽低，中间可以相差N多。这回测个blank的值，结果比我有细胞的值还高。。。不
知道是不是probe的问题。
一个probe大家大概用多久？几个月还是只要不坏，就一直用？我发现probe测出的读数
会越来越诡异。。。 |
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c******r
发帖数: 3778 | 40 为什么会GFP效率太低？用的什么promoter？
The simple way is to pre-label your rat cells before you inject them into
mice. There are variety of fluorescent probes you can use for long term cell
tracking. Just go to molecular probes web site, they have them pretty well
organized, so you can find the probe you need easily.
太低，没有用。
好一些吧 |
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b******n
发帖数: 492 | 41 最近需要设计probe for southern blotting to screen the founders， 有一些问题
想请教一下各位:
1）如果是用gene probe， 一般多长比较好？ 有人说越长越好， 可是大概是多少呢？
2） 如果用oligo probe， 50 bp 差不多吗？ 需要准备至少两个吗？
3） 我现在有好几个 enzyme only cut once in the transgene，但是其中好几个都可
能blocked or impaired by CG methylation, 是不是最好都avoid?
谢谢了！ |
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a****k
发帖数: 1130 | 42 30kD的话那你的RNA probe大概有100nt左右了吧？根据我自己的经验，100nt的RNA再加
上个200kD的蛋白，是很难跑进6%的acrylamide gel的，所以要是我的话一般会选择用
agarose gel。还有看起来你的sample是蛋白加的越多band就越强，我怀疑是不是你的
incubation buffer有RNase能degrade你的probe，然后很有可能你的蛋白本身能bind
probe，所以蛋白加多了也就保护了RNA |
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M****e
发帖数: 178 | 43 准备用NimbleGen-tiled microarrays做DNA methylation. 不知版上是否有前辈做过，
有几个关于数据分析的问题向请教一下。
1. 从机器上得到的原数据是否是每个probe的density？
2. Bioconductor 中有做ACME的package， 似乎可以得到significance (p value) for
each probe/region/promoter. 但这只是针对每个单独样品， 不能做样品之间的比较
。而且，这个p value能否当作quantitative value 来用， 还是选定cut-off p value
后只能得到yes/no for each probe?
3. Bioconductor 还有其他一些packages (Ringo, biomaRt, topGo), 好像也可以分析
DNA microarray data, 还能做样品间比较，但不知对NimbleGen-tiled microarrays
的data 是否适用？
如果有这方面的经验，能告知分析的大致流程，就再好不过。 非常感谢。 |
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m***o
发帖数: 272 | 44 在做conditional knockout。现在需要做southern筛选ES。测了几个probe都不好用，
老板着急，想找公司做。可是看了几个怎么需要我们提供probe，问题是如果我们有好
用的probe就不找公司做了。请问板上各位，有谁知道好的公司可以提供southern服务
，我们提供construct和电转的干细胞，也可以从干细胞里提好DNA。谢谢！ |
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L******e
发帖数: 679 | 45 Using TaqMan SNP genotyping assay to detect a SNP. The problem is that all
samples (blood genomic DNA) always showed the same "heterozygous" patten:
signals for WT and SNP are the same.
Does that mean the probes bind unspecifically? i tried different annealing
tep. at 60, 62 and 64 C degree, looks no much change.
The Tm of the primers: 55 C (forward) and 59 C (reverse)
The Tm of probes: 51 C (WT), 49 C (SNP)
PCR product: 90 bps
The primers and probes are designed by the ABI.
What can i do nex... 阅读全帖 |
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C******d
发帖数: 362 | 46 刚做real-time PCR，遇见了一个大问题，待高手解答，先谢谢了！
我的样品没有扩增曲线，但gel显示band is there.
PCR 反应溶液组分：
DNA 2 uL, 50-150 ng， 260/280 都在1.8-2.1之间
taqman probe: 2 pmol
primers: 4 pmol
master mix: 10 ul
总共： 20 uL
real-time PCR 仪器是：Stratagene Mx3000
昨天和合成probe的公司联系，说可能要加DMSO，今天就加了5%，结果同样是没有扩增
曲线？
primer和probe，都是按文献设计做的。莫非是仪器设置的问题？还是其他莫名原因？
跪谢！！！！ |
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z****u
发帖数: 1007 | 47 倒是想做LNA probe. 不过那一个probe得700多元，有点心疼的说。
有谁拿LNA probe做过ISH没？ |
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l**********1
发帖数: 5204 | 49 if that PCR mutation coincidencely happend on the key ponit or region of the
binding sites between that
probe and Genomic DNA sample, which is simialr to loss of docking site in
Western Blotting , then how can you expect both bind tightly unless p32 or
s35 etc isotope labelling can pick weak binging up but Dig labelling Can't
pick that kind of weak binging.
And after sub cloning that the slected white clone can keep your probe
almost no changed and inserted into that vector with LacZ gene of β-
... 阅读全帖 |
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B******o
发帖数: 496 | 50 Your information is too limited. What is your target? mRNA or small RNAs?
How about your probe, DNA or RNA?
For small RNAs, complementary DNA probe is more than enough.
For mRNAs, people used in vitro transcription to generate ~500nt DNA probe. |
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