p*****n
发帖数: 981 | 1 ☆─────────────────────────────────────☆
tataat (tataat) 于 (Sat Jan 20 23:34:03 2007) 提到:
我用SalI+NotI切一个片断（from a vector). 加了buffer 3和BSA. 结果发现之切了一
刀。于是作了个对照：+SalI only ; +NotI only; +SalI+NotI; +NdeI +NcoI(他们正
好在salI 和 NotI 的外侧，只差十几个bp).
结果：NdeI+NcoI: 切成了两个片断(3+0.4kb)，size都对。所以片断连进去了（pcr+
pgemt，0.4+3.0kb)
+SalI: linearized, only one band(3.4kb)
+NotI: linearized, only one band (3.4)
+SalI+NotI: linearied, only one band (3.4)
实在想不通，难道salI不能和notI混和吗？以前好像也这么干过，没问题的。
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b******r
发帖数: 111 | 2 Anyone has experience of inserting 9kb in a 8.4kb vector? 2 months was gone
,but I get nothing. Would
you like to take a look a this protocol and give me some suggestion ?
Thanks a lot
Purpose: Insert 9kb mouse sequence into vector pPNT6.
Vector pPNT6: Its map is at http://www.addgene.org/pgvec1?
f=c&identifier=11072&atqx=pnt6&cmd=findpl . I have inserted 0.9kb DNA in
this vector at XhoI/EcorI
double sites. I should have no problem in cloning short fragment.
Insert DNA is 9kb.I have cloned t... 阅读全帖 |
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h*******y
发帖数: 577 | 3 刷状态刚刷到
On February 24, 2015, we approved your Form I-140, Immigrant Petition for
Alien Worker , Receipt Number LIN150945XXXX. We will mail your approval
notice. Please follow the instructions in the notice. If you move, go to www
.uscis.gov/addresschange to give us your new mailing address.
---------------------------------------------------------------
timeline
2.01 开始准备要推荐信，写PL
2.18 材料寄出, USPS express mail, 60大洋
2.19 delivered
2.23 received email notie 显示RD date 为 2.19
2.24 received hardcopy n... 阅读全帖 |
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b********9
发帖数: 1061 | 4 最近一直在做cloning,一个3Kb, 一个2Kb，一个8Kb，连在一起，酶切为点：pstI,
HindIII, and NotI 怎么连也连不上，已经做了2个月了，哪位大虾帮帮忙？
3Kb: pstI and Not1
2Kb: notI and HindIII
8Kb: PstI and Hind III.
多谢。 |
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h***9
发帖数: 662 | 5 一个准备用来做knock-in老鼠的targeting vector（14kb),用invitrogen的kit做了
maxi-prep,然后用NotI linearize,我确定NotI是单一酶切位点,可是为什么出来这么多
大大小小的片断（4kb,2.5kb,1.5kb,100bp)？ |
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l**********1
发帖数: 5204 | 6 LZ please try below (I) to (IV) four strands overlap PCR
your target 7kp=2kp(I)+2kp(II)+2kp(III)+1kp(IV)
if you did not know downstream any conservative sequence information
your 3' RACE can use ligation-anchored PCR
就是用内切酶
//en.wikipedia.org/wiki/Restriction_enzyme
对应的人工引物锚定long mRNA 的3'端 做人工锚定引物
i.e. by NotI it should be inserted to 3'端 5'GC---GGCCGC
from
//en.wikipedia.org/wiki/Restriction_enzyme
Enzyme Source Recognition Sequence Cut
NotI Nocardia otitidis
5'GCGGCCGC
3'CGCCGG... 阅读全帖 |
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j**W
发帖数: 89 | 7 见新图并更正。
Sequencing was done with P2, which is within Exon 1. The resulting sequence
include region A (part of Exon 1 starting from P2 to before NotI site)
followed directly by region C (Intron 1 after the AscI site). NotI and AscI
are the enzymes used to put the construct together.
现在清楚了吗？ |
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l**********1
发帖数: 5204 | 8 GC contents of that P2 to AscI site if >65% DMSO 5% to buffer or try
Clontech GC rich kit
Cycle time 35 that is no problem
pls refer
GC-rich PCR concerns DNA templates with high GC content, which is defined as
the percentage of guanine-cytosine base pairs. When amplifying templates
with >60% GC content (i.e 5' ends of most mammalian cDNAs), the three
hydrogen bonds shared by GC pairs lead to enhanced thermostability, which is
problematic for un-optimized Taq polymerase systems.
http://www.clon... 阅读全帖 |
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u****n
发帖数: 7521 | 9 美国普通人非常痛恨过度的福利制度
主贴:
Tea Party 'tantrum' not good for nation
1:30 PM, Aug. 5, 2011 |
66Comments
http://www.stargazette.com/article/20110808/VIEWPOINTS03/108080
Thanks to the Tea Party. For them, it's my way or the highway — does that
remind you of your children's temper tantrums? Well, it worked!
Forget the poor, the elderly and the unemployed, while the wealthiest
continue to enjoy their quality of life at the expense of the rest of us.
God help us.
The Rev. and Mrs. Robert Adkins
Elmira
评论（被... 阅读全帖 |
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C*I
发帖数: 4736 | 10 ４０００亿美元的两房债券随时变现？ 别扯了。 两房债券是当年美国住房建设部长
伙同财政部长亲自到中国说服胡温在“做负责人大国的旗帜下”忽悠走的，压根就不是
从一级债券市场一点一点批发来的，而是给你签合同paper来的，上面的title明明白白
的写的是中华人民共和国。请问，你怎么拿去卖？ 又能卖给谁？ 金融危机那些年，两
房股票剩下几分钱，债券跌到和垃圾一个样子，4000亿美元估计连4000万都没有了。
中国当时还让美国政府保证这些债券安全，但美国参政两院一些议员明确表示: 保证什
么安全？投资就是有风险的，只想赚钱，不想承担风险的投资世界上没有。 让中国政
府直直的呛了一鼻子灰后不啃气了。
随后很长一段时期，一直到现在，不明真相的中国老百姓和一些所谓的傻逼“精英”与
愤青，时不时的不懂瞎逼干叫唤，说可以把这些债券卖了，瘫痪美国经济。 殊不知，
这些债券你压根就没有办法卖，要能卖，这不多少年了，你为何不卖一块二块的给全世
界人民看一看？ 更何况，今天中国外汇短缺，你卖点拿回来用啊！ 怎么还是分文不动？
至于说二级市场债券两房债券价格在市场利率暴低的时候相对涨了，那是不错。 可中
国买的... 阅读全帖 |
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g********d
发帖数: 19244 | 11 上月新闻指出，在过去的三十年里中国一汽集团一直非法剽窃其合作伙伴VW的引擎和变
速箱技术。本周早些时候，一汽正式回复了这些报道：一汽集团尚未从德国大众总部收
到任何关于技术剽窃的通知。
一汽发言人表示：如果VW真的认为我们抄了，VW应该先于我们私下商讨，而不是直接诉
诸媒体。当被记者问到为什么过了这么久才回应时，一汽说：VW还没给我们下战书，我
们觉得没有必要为这种无根据的指责回应。
这个流言最早出现于德国每日经济报Handelsblatt。该报道说VW的合作伙伴中国一汽抄
袭了EA111引擎和MQ200变速箱技术用于将出口给俄罗斯的Besturn B50（一汽奔腾）。
也有人怀疑这些流言源自于对VW与中国扩大合作不满的那些VW员工。
中国目前是大众最大的市场，今年上半年就售出130万辆大众汽车。大众与一汽和上汽
均有合作，并开始扩大产能以满足需求。VW和一汽在三个月前还签署了协议，两家将继
续在未来25年内合作。
http://autonews.gasgoo.com/china-news/faw-has-not-received-noti |
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j*****e
发帖数: 11116 | 12 pickup的时候，pickup的noti太多了，都是自己去的话估计要被打出来了 如果不管名
字就让别人去pickup
如果管名字 以后是不是reserve的时候都写mike，jerry啥的了 |
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s*****A
发帖数: 478 | 13 Dear Customer,
Thank you for your interest in purchasing the HP TouchPad. Due to the signi
ficant price reduction, we experienced overwhelming demand for the product a
nd are temporarily out of inventory. We apologize for the inconvenience, an
d unfortunately this means we are not able to complete your order at this ti
me. We have already canceled your order and started processing your credit.
In most cases your bank will post it to your account within 3 to 5 busines
s days.
For those of yo... 阅读全帖 |
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t*********u
发帖数: 26311 | 14 Dear Customer,
Thank you for your interest in purchasing the HP TouchPad. Due to the signi
ficant price reduction, we experienced overwhelming demand for the product a
nd are temporarily out of inventory. We apologize for the inconvenience, an
d unfortunately this means we are not able to complete your order at this ti
me. We have already canceled your order and started processing your credit.
In most cases your bank will post it to your account within 3 to 5 busines
s days.
For those of yo... 阅读全帖 |
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i**y
发帖数: 829 | 15 我知道部分解答：
H1B APPROVAL NOTICE：正本是1-2张彩色的纸张，I-797 NOTICE OF ACTION。其中NOTI
CE TYPE: APPROVAL NOTICE
H1B RECEIPT：就是I-797 收据 |
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v****t
发帖数: 338 | 16 Respected Dr Vicktt
I, A.Sriram, am currently a pre-final year student pursuing Bach
elor of Technology course at Indian Institute of Technology (IIT) Bhubaneswa
r, one of India's premier institutions for science and engineering.
I am looking for a research internship under your guidance(between 1st week
of May to 2nd week of July) which will be in keeping with my long-term obje
ctive of carrying out research in an academic position in my own country as
well as give me an opportunity... 阅读全帖 |
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F********n
发帖数: 244 | 18 The following works for me:
One nasal spray (NasalCrom) recommended by Mayo Clinic works very well, and
it is safe for pregnant women. But it takes a few days to build up the
protection.
I also bought two noti pots to rinse my nose several time with saline water.
One at home, on in office. Whenever I come back from outside, I rinse my
nose.
Air purifier helps a lot, we bought two, and let them run day and night.
Hope these will help. |
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s**8
发帖数: 67 | 19 --I think my landlord only
can charge my money from wall.
地板is good, I have taked picture. sink is blocked, so I complain
landlord, and landlord let worker repaired it , and take out middle sth by
repair worker.
Acturly my law saled me, court first notied time on march 17, but ld
vacation and arranged all sth, prepared to go to court that day, my law
calling ld that landlord cance that time case, and contine let my law argue
with us,so no resoult to argue , and we find anther legal service , ... 阅读全帖 |
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o*******0
发帖数: 577 | 20 4/4 ordered
4/6 shipping noti
4/13 got |
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b**********a
发帖数: 18 | 21 我想，看你们俩是谁为主邀请，被邀请的哪方的？两个都是工作的话最好是自己邀请自
己那方的，因为更容易让签证官相信你们之间的紧密联系。
>> 1. 需不需要我们的旧护照的复印件，
我认为是需要的。新护照是在美国换发的是吗？如此更是要了。而且，你旧护照有你
2008年回国记录吧？那是非常重要的证据。
>> 2.是否需要旧护照上的第一次来美国F1F2签证复印件
我认为也是要的。因为签证官很可能第一问题就是你女儿是怎么去美国的，这时候就可
能需要你第一次来美的材料了。
>> 3.是否需要旧I-94复印件
我建议多多宜善。不要吃我岳母第一次签证时因为我们材料准备不足的亏了。老人家当
时可伤心了。
>> 4.关于移民的复印件，是不是只提供绿卡复印件就行了，是否还需要提供过去办的
EAD
（工卡）复印件，I-485收据？
我也都附上了。我觉得关键是你如何呈现给签证官一个清晰的timeline，你的身份是一
步步怎么过来的。我不但把所有这些移民文件按时间顺序排好，还写了一封信给签证官
，把timeline用列表的形式写在上面，看起来比较方便。
至于你弄丢了的I-485收据（还是approval noti |
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s***m
发帖数: 6197 | 22 我还没hold options到expire过，没实践过
一下是我在CBOE网站上看到的
When can you be assigned on a short equity option position?
As an equity call or put option holder may exercise the contract at any time
before it expires, an equity option writer may be assigned an exercise noti
ce at anytime before expiration.
When do options expire?
Expiration day for equity and index options is the Saturday immediately foll
owing the third Friday of the expiration month until February 15, 2015. On a
nd after February 15, 2015, the... 阅读全帖 |
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d********8
发帖数: 2 | 23 我file 140时 使用了USPS EXPRESS MAIL,当时想挺重要的，邮局人问我要不要
SIGNATURE，我就说要。第二天TARCK说，NOTIE LEFT，要去邮局PICK UP或者要求
REDELIVERY. 移民局的人会去 PICK UP吗？还是我应给立刻要求REDELIVERY？ 多谢了
！ |
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u***s
发帖数: 591 | 24 ☆─────────────────────────────────────☆
cjsnyxz (eric) 于 (Thu Jun 22 18:42:09 2006) 提到:
已拿到加拿大移民签证，本人持H1B身份，欲与妻子(H4身份）最近开车登陆加拿大（温哥
华）。不知道回美风险大不大。可有大侠有类似经验？望告之， 多谢！！
☆─────────────────────────────────────☆
lkenlken (lken) 于 (Thu Jun 22 18:45:02 2006) 提到:
I did exactly the same thing last November.
Coming back to USA was very easy.
It was so easy that we actually went to Canada and come back again in this
April 2006 :)
You need following document to come back to USA:
1. H1B approval noti |
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J********i
发帖数: 50662 | 25 恭喜
在本版潜水很多日，学到不少东西， 现在终于收到卡了。
回报一下，也祝大家绿卡申请顺利。
广发包子，发完为止~~~
本人背景：
2009年US Ph.D.， 现在在做Research Scientist.
Journal Paper: 11, first author: 9, all good journals
Conference Paper: 16, most are first author (11/16)
Patent Application: 6, each of them has been applied for US patent and inter
national Patent
Review: 12 times for 3 journals
Citation: 50+
6 recommendation letters, 2 of them are independent. All are from big bulls.
Timeline:
12/28/09: 140,485,765,131 RD in TSC, non-PP
01/12/10: FP noti |
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O*O
发帖数: 2284 | 26 再等一周看看
at the USCIS web site: "On July 2, 2010, we mailed you a notice that we have
approved this I140 IMMIGRANT PETITION FOR ALIEN WORKER. Please follow any
instructions on the noti |
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a******t
发帖数: 416 | 27 Thanks, however, i am really not sure what will we get from the mail.
Did not get any approval notie before this. Jumped from initial review to
Card/Document production for I-140 case. |
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b*******e
发帖数: 24532 | 28 faint, you have signed up email noti ,right? |
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c******n
发帖数: 16403 | 29 30号寄出的， 刚刚收到email
On April 9, 2012, we mailed you a notice that we have approved this I140 IMM
IGRANT PETITION FOR ALIEN WORKER. Please follow any instructions on the noti
ce. If you move before you receive the notice, call customer service at 1-80
0-375-5283. |
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X***l
发帖数: 456 | 37 恭喜枫树，有审稿推荐我吧
IMM
noti
80 |
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Y***a
发帖数: 463 | 40 wow, gxgx! baozi!
IMM
noti
80 |
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F******1
发帖数: 304 | 43 GXGX, 发包子吧，是自己准备的？
IMM
noti
80 |
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e******r
发帖数: 9977 | 44 big con! background?
IMM
noti
80 |
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G*****h
发帖数: 2272 | 46 恭喜亚,介绍一下背景吧
IMM
noti
80 |
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