z**n 发帖数: 22303 | 1 http://www.spacetoys.com/proddetail.php?PARTNER=spaceguide&prod=spaceshuttleorbiter
New: Space Shuttle Orbiter 1:1 Model for $2,200,000 + pickup
SpaceToys.com offers a Space Shuttle Orbiter 1:1 Scale Replica Model for $2,
200,000. Although shipping is the responsibility of the purchaser, that's $
26,600,000 below the lowest total price we could for an actual NASA space
shuttle. This non-flying replica does not include an interior.
Posted 42 min ago More Gadgets |
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l********u 发帖数: 910 | 2 Failing student jailed for trying to bribe professor
Yang Li had a 0.177 air pistol with him when he went to meet his professor,
Bristol crown court heard. Photograph: PA
A failing student who offered his professor £5,000 in cash in an attempt to
pass his degree has been jailed for 12 months.
Yang Li, 26, had a loaded replica gun and the money in his pocket when he
went to meet Professor Andrew Graves at the University of Bath, Bristol
crown court heard.
Li had been given a mark of 37% in his di... 阅读全帖 |
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t***h 发帖数: 5601 | 3 http://www.marketplace.org/topics/world/chinas-hangover/china-r
By Rob Schmitz, Marketplace for Wednesday, July 3, 2013
The financial district of Yujiapu in the port city of Tianjin aims to be a
rival to Manhattan within ten years. The government has spent billions
building replicas of Rockefeller Center, Lincoln Center, and are building
China's largest high-speed rail station here. Economists question the
district's future.
A construction worker rests underneath a bulldozer amidst the rubble of... 阅读全帖 |
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s*****n 发帖数: 1794 | 4 民煮大国就是与众不同。
Cleveland police kill boy, 12, carrying a replica gun
A 12-year-old boy has been shot dead by police in the US city of Cleveland,
after carrying what turned out to be a replica gun in a playground.
Cleveland deputy police chief Ed Tomba said the boy was shot twice after
pulling the gun from the waistband of his trousers.
The boy did not make any verbal threats nor point the gun towards the
officers, Mr Tomba added.
http://www.bbc.com/news/world-us-canada-30172433 |
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发帖数: 1 | 6 12月26日 事实的真相
以前我们分析过,Augusta, GA 只是这个人体基地的一个点,而且我们也追踪到还有一
个点在加州。那么经过更多的分析,显示事实的真相可能是如此。这些地下通道是很久
以前就存在的,是为了战争或避难用的,后来在南北战争中,又充当奴隶逃跑到北方的
通道。那么在后来的后来,就被一些别有用心的人充当了非法活动的场地。而到了80年
代后,日本人就加入进来,把其改造成了一个军事,人体实验基地,成了报复美国的一
种手段。当然,我也强调历史和解,仇恨和解,但要在正确的认知,悔改下的和解。如
果一边在屠杀受害者,一边高谈和解,并千方百计掩盖事情真相的曝光,这不是悔改的
表现,最多是转移了屠杀的目标,我们不能原谅罪恶势力彼此勾结下的和解。根据分析
,这个基地的入口就在Deerfield, ILLinois. 世界每一个人都是这个基地的受害者,
都是这些大药厂的受害者,受害者以亿为单位来计算。你们很可能都在不知不觉中受到
了他们的伤害,请把信息传达到世界每一个角落,让我们一起携手和世界上最邪恶的势
力做顽强的斗争,他们现在还在屠杀无辜的受害者,每天在我家里杀人。请你们和我站
在... 阅读全帖 |
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k**L 发帖数: 3630 | 7 §5001. Penalties for entering into commerce of imitation firearms
(a) Acts prohibited
It shall be unlawful for any person to manufacture, enter into commerce,
ship, transport, or receive any toy, look-alike, or imitation firearm unless
such firearm contains, or has affixed to it, a marking approved by the
Secretary of Commerce, as provided in subsection (b) of this section.
(b) Distinctive marking or device; exception; waiver; adjustments and
changes
(1) Except as provided in paragraph (2) or (3... 阅读全帖 |
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n******0 发帖数: 232 | 8 自用了个前两天搞的三星本本,那个Microsoft Office 2010 Home and Student
Product Key Card 没啥用,就准备到医院里出掉降低点成本。医院里面当时有7-8个一
模一样的东东在买,查了下completed记录,也卖掉了很多。结果list了 两次都被
remove了,说违反了copyright!难道被人搞了?请老医生指教下。
Your listing has been removed.
We have removed your item because you have listed a CD key without the
original software. In order to avoid any concern of copyright issues on our
site, we do not allow any CD keys to be listed unless they come with the
original software; this is a precaution that we take in order t... 阅读全帖 |
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y****u 发帖数: 831 | 9 之前挂了耳机在ebay上卖(3种颜色,挂了三个listing,图不一样,文字标题除了颜色
描述都一样)
有买家买个一个,我也打印了label(还没寄)
昨晚收到ebay的邮件说
After reviewing your account, we've had to take the following action:
- Some or all of your listings may have been removed. A list of any removed
items will be listed further down in this email.
- We have credited any associated fees to your account.
We don't allow replicas, counterfeit items, or unauthorized copies to be
listed on eBay. Unauthorized copies may include things that are bootlegged,
illegally d... 阅读全帖 |
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z*****4 发帖数: 835 | 10 这种直接表明replica的也犯法吗?
ebay上各种replica啊 |
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j********x 发帖数: 2330 | 11 load balance要考虑很多具体的实现模式
我瞎说:
如果有一个central的dispatching handler,可以动态调整每种query往机器上的分布
同时query cache的存储空间和替换策略也可以向需求量大的商品类别倾斜
如果没有central的handler,那就p2p,那更好忽悠了。动态调整每种商品数据的
replica,如果query很多就放更多的replica在其他的peer上 |
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b*******d 发帖数: 750 | 12 来自主题: JobHunting版 - 贡献几个题 有几个题,觉得有点意思。
1. 证明任意一个7个node的无向graph,至少两个node拥有同样的degree。每个node不
能有指向自己的edge。
2. 一个array,size n,里面的数都属于range[0, n-1],有unknown的duplicates,找
到至少一个duplicate,constant space,linear time。
3. 设计一个persistence系统对外提供get(key), put(key,value)的功能,要求
每条数据3个replica,无single point of failure,能自动修复replica上的failure
。基本是distributed system的课堂讨论题。可以是p2p结构,也可以是server-worker
结构,但我给的是p2p。 |
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c******3 发帖数: 296 | 13 S3和oracle差了4小时,不象replica,更象是给第3方用的?
写个service-c吧。service-a把数据写入oracle之后立即(async)call service-c把同
样的数据写入S3。如果service-c fail,service-c就把同样数据从oracle中再读出来
写入S3。
service-b可以退休了。
换成MySQL也一样,如果MySQL to MySQL replica 太慢。service-c写入MySQL。 |
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m******0 发帖数: 222 | 14 请大家讨论一下吧!
我能想到的,在real面试中我可能会这么说:
1. 首先看用户和司机在app上都需要什么功能:
1)用户:设目的地、list附近的车,
2)司机:查看顺路乘客、查看附近乘客、自动报单供选择、直接分配单子
2. 根据需求,系统分为几层:
1)load balancer
2)web server
3)数据存储(包括user/driver的meta data,位置信息,历史订单与奖励信息、聊
天信息)
4)逻辑处理(user和driver的match,driver在接user的路上信息推送、奖励分配)
3. 具体每层系统的设计:
1* load balance:
1)按app的IP地址分配地理位置最近,或者latency最小的web server
2)在人口密集地(NY、BEIJING),选负载较少的server
2* web server层
1)nothing special
3* 数据储存和设计
1)user/driver的meta data存在key-value的cache中,如memcached,后面的
permanent ... 阅读全帖 |
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z*********n 发帖数: 94654 | 15 我没买名画replica (当然也许也是名画的replica我自己不知道而已,哈)
就是一些画得挺好看的风景画而已,我觉得挺好的,bad taste就bad taste
who pa who啊
Hang
usually
house |
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z****n 发帖数: 3189 | 16 看来是遇到同行了,老夫多说两句
1.自己写中间件,用一致性哈希算法解决负载均衡,sharding和replica的问题。
老夫倒没有研究过mongodb的sharding tool是不是有问题,看过dba操作过,没听他
有什么抱怨,但是老夫不喜欢被人控制,追求极致性能,所以自己写中间件没有用到
mongodb的sharding 和replica
2.Index的坑
老夫之前压力测试,一天100B个transaction , 70%写,30%随机删,3天之内index就
完蛋,没办法,b+tree做的index就是这样的了,这正是为什么老夫对他没有实现lsm算
法十分不满。但是有独特办法可以解决。
3.为什么用mongodb
当时是因为业务量飙升,需要紧急替代另外一套系统。老夫用nodejs写这个中间件做
service,居然2天写完可以开始测。一开始试过用mysql,发现性能还是mongodb好些。
设计负载量10k qps ,只需要10个instance就顶住了,性能让老夫喜出望外。nodejs的
mongodb驱动做得很可以,mysql的驱动做得比较渣。除了前期的坑,老... 阅读全帖 |
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b******d 发帖数: 9694 | 17 作为狮子队多年的忠实球迷很幸运的被选中参加明晚比赛的“Fan Appreciation
Tunnel”活动, 具体活动就是在比赛前球员从Lockerroom出来的时候和他们握手致意
,零距离和他们接触,然后和其他幸运球迷到球场中央列阵参加唱国歌仪式。 因为比
赛全国转播, 所以你的亲朋好友同事极有可能在电视上看到你。还有就是活动开始之前狮子队应该会给每人一件球衣。Hopefully it is the authentic one, not the
replica one. The authentic one costs around $150-200 from many team websites
and the replica one is usually worth around $30-60.然后就是回到座位静心看比
赛。比赛是晚上8点钟开始, 但是为了参加这一系列活动我们需要6:45pm到球场。
球票我们已经买好了,本来是我们一家四口要去, 但是老婆临时有事去不了。因为参
加这个活动的人员球队都是提前登记好的, 这么好的机会觉得浪费了可惜, 所以特意
征一女球迷前往, 代价就是... 阅读全帖 |
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c*******e 发帖数: 514 | 18 2009 Lamborghini Murcielago Replica
keyword REPLICA |
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s*********5 发帖数: 5637 | 19 原文链接:(点击后有每个故事的详情)令人难过的是很多故事都是与校园有关的。某
些学校的教师校长甚至都有配备保安。我知道我们孩子的学校里每个教室都配有对讲机
,有突发事件时门也会自动锁起来,孩子每过一段时间就会防火,防震,防暴演练一次
,就是要防范类似事件的发生。这是一个怎样的时代呀。。
http://delmar-carmelvalley.patch.com/groups/police-and-fire/p/1
We looked at stories from across America in the year since the shootings at
Sandy Hook School to try to illustrate the scale of the problem.
This article is a work of aggregation building on the reporting of scores of
Patch reporters across more than 900 sites. It was written by John Ness an... 阅读全帖 |
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g*******4 发帖数: 155 | 20 背心?你说的是球衣吧?
球衣分Replica, Swingman, Authentic 三种,质地和价格差很远。
各种球队还有不同的球衣,主场客场以及备用,还有很多复古版,纪念版,甚至特定版
(例如“拉丁之夜”系列)
自己上nba.com/shop去看吧。我自己买到最便宜的球衣,10美元,弱队过气球星超大的
Replica。最贵的Jordan Authentic 复古纪念版的官网要400美元,差的太多了 |
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j****3 发帖数: 228 | 21 NBA的球衣价格区间很大的,便宜的Replica $10-$50,贵的Authentic的官方正品要$
100-$500, KOBE或Lebron的球衣价格是无名小卒的好几倍。如果关系一般的话买件
Replica就可以了。 |
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l******l 发帖数: 2651 | 22 来自主题: SanFrancisco版 - 山寨福音 多好的例子.
国内跟老牌帝国主义比还是太嫩
Next one will be Disney
http://insidemovies.ew.com/2011/07/27/george-lucas-loses-legal-
George Lucas loses legal battle over 'Star Wars' Stormtrooper helmets in U.K.
by Lanford Beard
Categories: George Lucas, Sci-Fi, Star Wars
Comments 32 Add comment.Reuters reports Britain’s high court decided today
that George Lucas did not have a case for copyright infringement against
Star Wars prop designer Andrew Ainsworth… at least in the U.K.
Ainsworth, 62, kept the original molds a... 阅读全帖 |
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a****0 发帖数: 209 | 23 replica一般材质比较低级,胶印很快就坏了,
swingman版型nike曾经做的很好,内里的刺绣很扎人,绣上的字是花纹是印的,会退色,
authentic都是类似于球员身上穿的材料,刺绣也是多层,不会掉色,纪念价值最高,
和球员穿的材质和做工都只能说是接近,还是有差距的,
authentic打球休闲都可以,replica也是(但是坏得快),swingman里面最好套个Tee
至于充门面,还得是Game issued,Game used之类的,authentic不够级别的 |
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n******u 发帖数: 154 | 24 which one is better:
Jeremy Lin Adult Red Revolution 30 Replica Jersey or Jeremy Lin Adult
Alternate Revolution 30 Replica Jersey? |
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w****a 发帖数: 1623 | 25 http://www.rentalboatcharters.com/13038
CAN WE KEEP OUR FISH?
We practice catch and release as part of our conservation effort. If you
would like to have a fish mounted, we have a taxidermist who can do a
replica mount, and ship it to you when it is completed. A replica looks like
the real thing, and often lasts longer. If you want to eat fish, there are
other species that are better eating than bass, like catfish, which we will
allow you to keep. |
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b******d 发帖数: 9694 | 26 作为狮子队多年的忠实球迷很幸运的被选中参加明晚比赛的“Fan Appreciation
Tunnel”活动, 具体活动就是在比赛前球员从Lockerroom出来的时候和他们握手致意
,零距离和他们接触,然后和其他幸运球迷到球场中央列阵参加唱国歌仪式。 因为比
赛全国转播, 所以你的亲朋好友同事极有可能在电视上看到你。还有就是活动开始之前狮子队应该会给每人一件球衣。Hopefully it is the authentic one, not the
replica one. The authentic one costs around $150-200 from many team websites
and the replica one is usually worth around $30-60.然后就是回到座位静心看比
赛。比赛是晚上8点钟开始, 但是为了参加这一系列活动我们需要6:45pm到球场。
球票我们已经买好了,本来是我们一家四口要去, 但是老婆临时有事去不了。因为参
加这个活动的人员球队都是提前登记好的, 这么好的机会觉得浪费了可惜, 所以特意
征一女球迷前往, 代价就是... 阅读全帖 |
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s****h 发帖数: 3979 | 27 衣服穿大号的,Replica Football Jersey是不是应该买小号?
N年前网上买了件中号的Replica Football Jersey,大得不行。
这两年胖了些,但也要在里面衬件长袖才勉强能穿出门。 |
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t******8 发帖数: 2803 | 28 wiki上的:
Federal law in the United States requires that a 6 mm (0.24 in) orange tip
to be present on all "toy guns" (including airsoft replicas) while being
transported within or imported into the United States. These brightly
colored tips show the difference between real and replica firearms, which
helps to ensure safety.[15] However, when playing on a private field, no
orange tip is needed. In strict technicality, the federal regulations do
not specifically require the owner to keep the muzzle ... 阅读全帖 |
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g*****t 发帖数: 776 | 29 。。。。。
。。。。。
然后那位居民很疑惑地问我,but...I thought you were a Chinese too...
我说,Yes, I am. But I'm a good Chinese who upholds the laws. Look at your
dog, he doesn't even bark at me.
正说着,那位居民指着一个推着破自行车的亚洲人说,there's one coming. 我一看,
这位头上缠着红布,车把上挂一把捡来的玩Call of Duty用的仿AR15,后面车架上驮着
两个装满空罐头空塑料瓶的垃圾袋。。。。
我说,I know him. He has been street checked by us many many times and
stripped searched twice. I think his name was "Hetzer MAO". He's a weirdo.
We found a shit stained Chairman Mao's photo between his crack cheeks... 阅读全帖 |
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e********e 发帖数: 435 | 30 "Our Promise
We then inspect every nook and cranny and purchase the same materials to
ensure that the products we manufacture are virtually indistinguishable from
the designer originals in every way. There are cheaper replicas on the
market, but there are no better replicas available anywhere." |
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S*******7 发帖数: 3 | 31 www.leatherbrandbags.com aims to be best replica bags and wallets supplier
in the world. We sell the top grade replica handbags, all of them are
genuine leather handbags and wallets,,while the price is much competive than
the authentic designers handbags you want to pay. All series of products
are used by Italy imported cowskin, sheepskin, and France imported skin mix
purpose-built hardware to be refined,the hottest brands you can find like
Louis Vuitton, Marc Jacobs, Prada, Gucci, Miumiu, Fendi |
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S*******7 发帖数: 3 | 32 www.leatherbrandbags.com aims to be best replica bags and wallets supplier
in the world. We sell the top grade replica handbags, all of them are
genuine leather handbags and wallets,,while the price is much competive than
the authentic designers handbags you want to pay. All series of products
are used by Italy imported cowskin, sheepskin, and France imported skin mix
purpose-built hardware to be refined,the hottest brands you can find like
Louis Vuitton, Marc Jacobs, Prada, Gucci, Miumiu, Fendi |
|
l*******g 发帖数: 4894 | 33 琴其实就是块木头,gibson的59年的木头现在只有max一个人有。以前用的胶现在也不
用了。以前用的漆现在也不用了。那把81年的是准备开始reissue的时候大家纷纷模仿
的一把。所以纪念价值很大,我昨天绝对是头晕了,现在后悔到了极点。准确的说我标
完失败以后一分钟没说出话来,当时真是后悔到肠子都绿了。没有听kurtz的教诲阿,
不过后来觉得可能是被人作弊了,因为我用手机看到的最后bid的这个人就是之前一直
停留在2450的人,但是2450这个价格在list也没有,不过总而言之,那把琴没了。
max前段时间我和他通过信,他自己做的59 replica,要价$26,000. 如果要别的款,同
样的木头,同样的胶同样的声音(pickup),最便宜的一把$7000,还有2把$10,000的
。其实我当时想如果他的59 replica在$15,000左右,我可能准备攒一年钱买一把。
ebay上他的琴used卖19,000. 毕竟这个人现在50岁了,然后他用的木头现在已经绝
迹了。做一把少一把。
再后来我想26,000=一辆车的价格,一个普通的apt的首付,无数顿饭和旅游的钱,我为
了一把琴值 |
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d********0 发帖数: 5142 | 34 ☆─────────────────────────────────────☆
mercuriusl (Mercurius) 于 (Thu Dec 16 00:40:36 2010, 美东) 提到:
ghetto
☆─────────────────────────────────────☆
Minsco (Love 胖P!) 于 (Thu Dec 16 01:25:35 2010, 美东) 提到:
pirate or knock-off
☆─────────────────────────────────────☆
deadpig (死猪) 于 (Thu Dec 16 01:27:48 2010, 美东) 提到:
shanzai
☆─────────────────────────────────────☆
monokeros (独角兽) 于 (Thu Dec 16 01:30:21 2010, 美东) 提到:
pirated
☆─────────────────────────────────────☆
ivanhuang881 (iv... 阅读全帖 |
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i****a 发帖数: 36252 | 35 Top 15 Surprises in Bill Gates' New Mansion
15 Sign over king-sized bed declares, "Use of the Words 'micro' and 'soft'
strictly prohibited while in bedroom."
14 No paintings, but live artists actually hanging on the wall.
13 Drawbridge is raised and lowered by hand.
12 Entire state of Rhode Island relocated to east wing.
11 Raises guinea pigs in his bedroom for extra cash.
10 No toilet paper, but handy stack of $100 bills.
9 Secret passage in library leads to Nerdcave where Bill keeps the
Nerdmo... 阅读全帖 |
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z**n 发帖数: 22303 | 36 【 以下文字转载自 Military 讨论区 】
发信人: zoun (zoun), 信区: Military
标 题: 军迷们圣诞礼物的一个选择
发信站: BBS 未名空间站 (Thu Nov 18 10:48:11 2010, 美东)
http://www.spacetoys.com/proddetail.php?PARTNER=spaceguide&prod=spaceshuttleorbiter
New: Space Shuttle Orbiter 1:1 Model for $2,200,000 + pickup
SpaceToys.com offers a Space Shuttle Orbiter 1:1 Scale Replica Model for $2,
200,000. Although shipping is the responsibility of the purchaser, that's $
26,600,000 below the lowest total price we could for an actual NASA space
shuttle... 阅读全帖 |
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M********n 发帖数: 4650 | 37 abundance 范围广了,理解也可以不同。甚至replica 故意回避具体数字也可能。
Replica是机器啊,若不是刻意隐瞒,应该是趋向于精确的。再说机器擅长的就是复制
东西。
不过我的感觉,ancient这个种族,应该是反映了西方人看待古代东方文明的态度,他
们对这种追求精神层次的平和和超然于善恶之外的修为是不以为然的,还是觉得需要他
们所认为的humanity来擦屁股。西人就是争强好胜,崇拜技术远胜过精神追求。 |
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o*****h 发帖数: 2082 | 38 replica也分等级了,最高等级的应该是瑞士产的吧,就是厂家自己replicate自己的,
其次是意大利货,最后是深圳货(貌似现在又有了越南货?),价格分布在几千到几十
美元
你们列的这些牌子,天坛的红桥市场几乎都能找到,想不被人BS为replica,很难
如果真想买个走得准的,20美元的waveceptor,300万年误差不超过1秒
上次回去,我家邻居的大娘买菜用的袋子是几可乱真的LV,还抱怨说便宜没好货,不结实
神马都是浮云 |
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H*****D 发帖数: 121 | 39 add a dock/port replica,
then use the DVI-to-hdmi adaptor and hdmi cable
laptop->dock/port replica DVI-> DVI to HDMI adaptor->HDMI cable->TV HDMI |
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k****i 发帖数: 1072 | 40 replication factor 3和quorum写,写了两个replicas然后return write failure了怎
么办?Retry和write-ahead log有什么问题?改为写所有replicas原理上和魏老师的有
什么区别?
所谓
他的 |
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g*****g 发帖数: 34805 | 41 Take a page from DB cluster design. You use a replica for long running job.
Updates are logged, batched and asynchronous.
If your objects are in memory, logging does you no good, but you can still
batch and update async. And if memory is a concern, who says replica can't
be in a different machine. |
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z****e 发帖数: 54598 | 42
distributed file system应该不是通过journal
错误毕竟是小概率事件,一个node的错误,可能性比如是1%
那么两个nodes,也就是replica = 2同时丢掉这个数据的可能性就是
1%^2,万分之一,如果设置replica = 3的话,那就是1%^3,就更低了
用这种方式来排错,注意设置逻辑version,做一个eventually consistent就好了 |
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T********i 发帖数: 2416 | 43 你有没有试试MongoDB?
貌似你这个是强调读性能。
MongoDB Replica Sets貌似可行。
By default, clients reads from a replica set’s primary; however, clients
can specify a read preference to direct read operations to other members.
For example, clients can configure read preferences to read from secondaries
or from nearest member to:
reduce latency in multi-data-center deployments,
improve read throughput by distributing high read-volumes (relative to write
volume),
perform backup operations, and/or
allow reads during failove... 阅读全帖 |
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R*****w 发帖数: 447 | 44 学习了,
请问怎么知道plasmid 是否有SV40 replica site? pcDNA3.1 有SV40 replica site吗?
protein make the cells grow slower, the untransfected cell will out-grow
the transfected ones in a few days. Assuming the 293T cells replicate every
24 hours, then in three days, the 10% untransfected cells will become 8
times of their original number and therefore will reach almost 80% of the
total cell population. |
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l***d 发帖数: 1828 | 45 http://www.livescience.com/culture/brain-3d-map-copy-cat-101115.html
Chinese team's brain imaging device has come under question from developers
of a U.S. device who say it's a near duplicate of theirs, LiveScience has
learned. An article on the Chinese device was published in the prestigious
journal Science, and the U.S. researchers are preparing a formal letter to
the journal in response.
According to the report appearing in the Nov. 4 online edition of Science,
the Chinese imaging device used... 阅读全帖 |
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s******y 发帖数: 28562 | 46 这个实验不是这么做的。因为大部分情况下面,不同的primer 所用的
PCR condition 不一样,所以不推荐在同一个cycle 里运行。
你应该是个菜鸟吧?那就不要乱改protocol 了。
这个实验应该这么做:
1. Primer gene G premixed, then add template control and template treated separately, with 3 replica .
2. Primer GAPDH premixed, then add template control and template treated separately, with 3 replica
所以要跑两次PCR. |
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c******r 发帖数: 3778 | 47 应该可以做multiplex reaction
GAPDH是可以做所谓的internal control的。就是说,在同一个pcr中,同时run两个甚
至两个targets,其中一个是control,另外一个是要测的基因。
这个internal control的目的有两个,一个是quality control,就是保证这个
reaction没有忘掉任何东西,没有加错任何东西等等。另外一个目的是loading
control,assumption就是两个sample中的GAPDH是不变的,换句话说,要测量的是在同
样量的GAPDH下,要测的基因在处理前和处理后有无变化。
为了达到这个目的,最好是run multiplex。这样才能保证测到的GAPDH和G之间是来自
确定的同一个loading量。
另外一个run multiplex的原因是统计的原因。如果分开run GAPDH和G,那么
triplicate之后会得到三个G的量,和三个GAPDH的量,而这六个量之间没有一一对应的
关系。就是说不能用G1/GAPDH1,G2/GAPDH2。。。。而只能用G的平均值/GAPDH的平均
值。但是... 阅读全帖 |
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Z******5 发帖数: 435 | 48 我设计引物的时候都会设计成同一个退火温度的。一次PCR搞定,这样比较方便呢。
separately, with 3 replica .
separately, with 3 replica |
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j*******u 发帖数: 81 | 49 这个楼主其实一直悟的就是这种东西,正经东西没想明白过
你看他发的这些帖子,biological replica和technical replica的分别一直就没搞清楚
这种人做的东西他就算不是有意造假我觉得也是不太可信的,因为他自己也不知道是真
是假
自己
发信人: RodinSophie (RodinSophie), 信区: Biology
标 题: 问个细胞实验重复的问题
发信站: BBS 未名空间站 (Wed Apr 6 11:09:25 2016, 美东)
打算做RNAseq,是不是要重复两次。
重复应该从什么时候开始算起?
处理完一碟细胞,分成两份,分别提RNA,算重复?
还是分成两碟,分别处理,算重复?
其实就算分成两碟,其实是从一管冻存的细胞复苏来的啊,还不是一样。
感觉这种重复没有太大意义啊。 |
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