T****i
发帖数: 15191 | 1 Roche 的一个kit。
Southern 比northern 要求低。不过你要是screen ES cells，还是用同位素保险。 |
|
d****d
发帖数: 214 | 2 我用过Roche的kit做Southern。曝光就象Western一样，不到10分钟就搞定，用同位素
压片则可能要几天的时间。DIG的缺点是如果掌握不好，背景会比较高。 |
|
F******p
发帖数: 2099 | 3 大家能不能推荐一个重组ES cell southern的kit? 一些专家告诉我DIG在ES cell上敏
感度不够。 |
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l**********1
发帖数: 5204 | 4 You can try AlkPhos Direct Labelling with GE RPN3690 kit and CDP-Star
Reference:
//www.ncbi.nlm.nih.gov/pubmed/21375737
citation:
Twenty-nine colonies were identified and expanded, and colonies were
subsequently genotyped by non-radioactive Southern blot (GE RPN3690 kit
with CDP-Star) with a 500-bp probe 5’ to the targeting vector and following
the manufacturer’s directions for labeling,
Manufacturer’s Protocol link:
HTTPS//www.gelifesciences.com/gehcls_images/GELS/Related%20Content/Files/
1314... 阅读全帖 |
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s******a
发帖数: 472 | 5 Has anyone been successful with Amersham AlkPhos Direct Labeling Kit to
perform mini-southern?
I mean ES clones are grown in 96-well plates.(You don't have many cells)
Or do I really have to go radioactivity?
The problem is that I can not perform radioactivity related experiments
without licence. |
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s******a
发帖数: 472 | 6 I used Amersham Direct Labeling Kit for my southern blot, and the protocol
is basically the kit manual. However, I have now a reproducible problem. As
shown in the blot, I expect a specific band in digested genomic DNA, but all
I have is a big smear (inside which there seems to be a band higher than my
target band).
I might have at least two problems:
1, the probe is bad;
2, the digestion is incomplete;
I kindly ask you to give me some advice regarding trouble shooting and how
should I proceed.
... 阅读全帖 |
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b*****o
发帖数: 150 | 7 我觉得是这样的
Genomic DNA如果不干净 在酶切体系中是很粘的一个球 没有彻底溶解 pacI(切8bp位点
很少) 很有可能没法进入球的内部 导致不完全消化DNA
对于一般6bp的酶 因为位点比较多 酶一边切割表面的DNA 一边会帮助DNA溶解 所以酶
可以渗透到酶的内部 切割会很完全 不会有类似的问题
我建议你用pacI和一个不影响这个位点pattern的6bp酶来一起切 就不会有这个问题了
PS 我最多的时候一年做50个southern p32的哦 呵呵 不堪回首啊 |
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c********u
发帖数: 250 | 8 transgenic mouse southern blot, 做了几个星期了还没出来,首要的问题如图，坑爹
的genomic DNA 用HindIII切了24个小时，60v 跑了6个小时，就给我跑成这副鬼样子，
大家提DNA， 溶DNA，切，跑胶有什么trick没？ 还有一个问题是，我的目标条带有
10kb，DIG probe, 我老板说10kb太大了，2-5kb最好，那我这种情况有可能成不？
真心求教，酌情发包子，谢谢。 |
|
s********r
发帖数: 312 | 9 southern这种实验耗时长，步骤多，加上DIG本身信号就比同位素弱，真心坑爹。新手
的话能搞的死去活来，我前段时间自己也是试了两次都没成功，实在不想伺候了，最后
以一顿buffet的代价请lab里一老手postdoc做的。。 |
|
a******8
发帖数: 56 | 10 You can see your wells has strong signal. DNA digestion is not complete.
HindIII is NOT a good enzyme for southern. try other enzyme and then trouble
shoot other conditions. Good luck.
Your marker is good, I guess that you also labeled a small amount of marker,
which is good. |
|
f*******d
发帖数: 92 | 11 这个是明显的酶切没有切开阿。DNA不干净应该是根本问题。 southern blot不是什么
很复杂的试验，一般只要DNA是好的，都能出结果。 |
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a******n
发帖数: 392 | 12 Southern做出来结果是一片smear，看不出条带。有意思的是没有标记的分子量Marker
倒是显示出条带来了。最有可能是什么问题？探针的特异性出问题了？我是用DIG PCR
标记的500bp的探针。探针电泳条带挺清晰的。 |
|
i*********0
发帖数: 915 | 13 你的probe是从载体里面切的么？
我以前做Southern也遇到这样的问题。我的probe也能在marker上产生signal，但是在
我的样品上是特异的带。
我估计是切下来的探针也带了一点plasimd的序列，和marker有cross reaction。
不过你在样品上看到smear带，可能是washing的不彻底，或者探针不特异。 |
|
s******a
发帖数: 472 | 14 非放射性方法确实信噪比差啊
记得哪里看到，说是southern发展了这么多年，虽然各公司如何吹嘘自己的kit，
DIG/AlkPhos等等
还是比不过放射性方法 |
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a******n
发帖数: 392 | 15 Nuclease残留是个可能的原因, 但是用southern作筛选克隆的时候，都是用粗提的
genomic DNA的，就是用proteinase K处理过夜，然后异丙醇沉淀。怎么尽量除去
nuclease呢？ |
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b****r
发帖数: 17995 | 16 人都说了试了好久都不行，你就肯定比人家聪明？
做实验不要想着cut corner，southern也不是那么可怕的东西。实验有比什么都重要
good |
|
r***e
发帖数: 2539 | 17 要不用roche的那个AP标记，做southern?
灵敏度比isotope略差，做genotyping应该够了，至少比较安全。 |
|
n********b
发帖数: 27 | 18 There is a position opening for a Principal Lab Assistant in a new Genomics
Center of a University located in southern California. The position is
supported by hard money with good benefit plans. Please send your resume
and email to [email protected]
/* */, if you are interested. Thanks!
------------
Principal Lab Assistant
Center for Genomics
Job descriptions:
1. Performs bench work related to research projects using different genomic
technologies including next-generation sequencing (N... 阅读全帖 |
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m*****a
发帖数: 658 | 19 Hi,
is it a good choice to study chemical engineering in the University of
Southern California ?
easy to find a job in the future?
Thanks a lot |
|
g*********i
发帖数: 89 | 20 Hi all,
We're looking for good RFIC design engineers, preferablly with power
amplifier and CMOS switch experience.
We're a startup in southern california.
I'm the hiring manager.
So you can send me yor resume directly if you're interested.
You're welcome to leave a message in MITBBS if you simply want to know more
about the opening.
Thanks. |
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t*****0
发帖数: 13 | 21 老板希望招新学生，有funding支持，有兴趣可以联系他。下面是他的主页：
http://lyle.smu.edu/~jhc/
学校：Southern Methodist University
地方: Dallas, Texas
方向：模拟射频集成电路方向，现在我们实验室做的几个方向有:High speed SerDes,
RF, AD/DA. |
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h*****r
发帖数: 81 | 22 The Water Quality Research and Development Center at the Southern Nevada
Water Authority (SNWA; www.snwa.com) is actively recruiting applicants for a
paid graduate internship in Henderson, Nevada. The Graduate Intern would
work at the SNWA R&D lab while enrolled in a graduate level academic program
. The duration of this internship is tentatively for 12 months with a
possible extension up to 18 months.
PRIMARY RESPONSIBILITIES The Intern will provide field assistance and
support for current proj... 阅读全帖 |
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r****x
发帖数: 1250 | 23 Mid sized biopharm in southern cali, looking for entry level formulatioin
scientist, background with liposome will be a huge plus.
Also looking for research associate (I, II or senior), BS/MS in
pharmaceutical sciences, chemistry, biology or related area. Pharm. industry
experience is a plus. Knowledge and exp with GLP/GMP regulations.
Experience with commonly used analytical techniques. Exp with material
testing (Instron, Zwick or similar), particle sizing is strongly preferred.
Pls. contact me |
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w*******y
发帖数: 60932 | 24 If you are planning on joining the Southern California Auto Club (AAA), you
can save $18 by going to the site Link:
http://www.aaa.com/joinnow
instead of the main site Link:
http://www.aaa.com/
. The membership is the same ($48) but the "new member admission" is $2.00
instead of $20.00. You also get a free travel mug.
Please don't turn this thread into the pros and cons of AAA membership!!! |
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w*******y
发帖数: 60932 | 25 Bus service around Southern California for $1 for booking in advance and
online.
Link:
http://www.crucero-usa.com/
Price range from $1 and up depending on destinations and advance purchase.
Just recently took a trip from Los Angeles to San Diego on the bus . Pay
only a $1 for one way trip. There was wifi on the bus but it was really
spotty.. so probably not worth bringing your laptop.
Looks like the company is an affiliate of Greyhound, so not the best service
and not the best bus stop locations... 阅读全帖 |
|
w*******y
发帖数: 60932 | 26 Groupon:
http://www.groupon.com/deals/kyochon-chicken?utm_campaign=kyoch
has an offer of $7 for $15 Worth of Korean Fried Chicken and Drinks at
Kyochon.
Limited to these Southern California locations:
Mid-Wilshire (Los Angeles)
Westfield Mall (Culver City)
Cerritos Mall (Cerritos)
Glendale Galleria (Glendale)
The Fine Print:
Expires Jul 7, 2011
Limit 1 per person, may buy 1 additional as a gift. Limit 1 per order. Limit
1 per visit. Dine-in and carryout only. Must use in 1 visit, no cash back.... 阅读全帖 |
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w*******y
发帖数: 60932 | 27 DESCRIPTION
Catalina Island: home of friendly orange Garibaldi, majestic kelp forests,
and the kind of placid peace and quiet you cant find anywhere else in
Southern California. It is an island, however, and getting there either
takes a mighty pair of swimming arms or a quick boat ride across the channel
. But with this deal you can get round trip fare to Catalina Island for just
$34! Departing from Newport Beach, the Catalina Flyer is the largest
passenger-carrying catamaran on the west coast. ... 阅读全帖 |
|
w*******y
发帖数: 60932 | 28 Southern LivingStaffordshire Bedding Coordinates 85% off with Kohl's
Charge + FS
Full/Queen Quilt
original $129.99
now $25.99
w/30% off code $18.19
Queen Comforter set
original $289.99
now $57.99
w/30% off code $40.59
King Comforter set
original $319.99
now $63.99
w/30% off code $44.79
Decorative pillows/Euro Shams and Drapes also available
Link:
http://www.kohls.com/kohlsStore/clearance/homestore1/bedding/PR Living Staffordshire Bedding Coordinates.jsp
Free Ship Code - FUN2BMVC
30% off code - L... 阅读全帖 |
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w*******y
发帖数: 60932 | 29 Southern States has a 20lb propane tank refill for $12.99 with coupon until
July 5th. This is a great deal since they actually fill it to 17 instead of
the 15 that the exchanges do and it is cheaper.
Here is the link on their site: Link:
http://www.southernstates.com/promotions/Independence-Day-2011. States Site Members&utm_campaign=d84fec300f-july4_blast2_06272011&utm_medium=email
Attaching a pdf of the coupon in case the link craps out.
|
|
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a******e
发帖数: 6689 | 31 【 以下文字转载自 LeisureTime 讨论区 】
发信人: aixiaoke (流浪者的乡愁), 信区: LeisureTime
标 题: 走或留，这是个问题
发信站: BBS 未名空间站 (Tue Dec 4 16:54:22 2012, 美东)
我非常喜欢美国年轻导演贝赫·泽特林 （Benh Zeitlin）的处女长片《南国野兽
》（Beasts of the Southern Wild）。这当然与我对新奥尔良的感情脱不开干系，但这
不过是一个起点。泽特林的镜头语言充满了质朴的张力，在情节与情绪的游走间他拍出
了美国南方腹地的湿度、味道与精神，这实在了不起。
电影最美的镜头之一是六岁的野孩子Hushpuppy在风暴来袭前与父亲Wink和“浴盆
”（Bathtub）地区的居民们欢庆南方节日。小小的Hushpuppy拿着璀璨的烟花在夏天湿
漉漉的海风中奔跑，活动房车里大人们疯狂地喝酒、吃虾、吃蟹。那是今朝有酒今朝醉
的南方，大快活里的温柔乡。
但风暴终于还是要来。泽特林为这场风暴加入了神秘的“原牛”——200万
年前牛的... 阅读全帖 |
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s*****t
发帖数: 1994 | 32 Southern Comet
Credit & Copyright: Noel Munford (Palmerston North Astronomical Society, New
Zealand)
Explanation: After a remarkable performance in the northern hemisphere, the
brightest comet in decades is now showing off in the south. Recorded during
evening twilight on January 17, this view features the bright coma and
gorgeous, sweeping tail of Comet McNaught (c/2006 P1) over Lake Horowhenua
in Levin, a small town on New Zealand's North Island. Astronomer Noel
Munford reports that the five s |
|
c*********e
发帖数: 13906 | 33 Worldwide M1+ Earthquakes, Past 7 Days
https://explore.data.gov/Geography-and-Environment/Worldwide-M1-Earthquakes-
Past-7-Days/7tag-iwnu
Src,Eqid,Version,Datetime,Lat,Lon,Magnitude,Depth,NST,Region
nn,00388838,9,"Tuesday, September 18, 2012 11:49:45 UTC",36.0628,-116.1913,1
.1,4.90,15,"Central California"
nc,71844125,1,"Tuesday, September 18, 2012 11:24:56 UTC",38.8212,-122.7993,1
.3,3.00,26,"Northern California"
hv,60400351,2,"Tuesday, September 18, 2012 10:48:50 UTC",19.0077,-155.4210,2
.9,44... 阅读全帖 |
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T******g
发帖数: 21328 | 34 不明白你想说什么，那个是世界排名，当然所有国家都有
你不是大嘴一说，美国超过世界一半以上的严重犯罪
你不会读数字，我只好给你贴出来了
UNODC murder rates most recent year (full table here)
Country Rate Count Region Subregion
Burundi 21.7 1,726 Africa Eastern Africa
Comoros 12.2 85 Africa Eastern Africa
Djibouti 3.4 29 Africa Eastern Africa
Eritrea 17.8 879 Africa Eastern Africa
Ethiopia 22.5 20,239 Africa Eastern Africa
Kenya 20.1 7,733 Africa Eastern Africa
Madagascar 8.1 ... 阅读全帖 |
|
s******r
发帖数: 5309 | 35 president privately called Sessions a “traitor,” saying: “This guy is
mentally retarded. He’s this dumb Southerner . . . He couldn’t
even be a one-person country lawyer down in Alabama.”
南方红脖子突然发现自己的偶像纽约人疮破私底下认为南方人是智障，草。。。。
Southern Republican senators defended Jeff Sessions after an explosive new
book by Bob Woodward recounted how President Trump called his attorney
general a “dumb Southerner” and mocked his accent.
In the forthcoming chronicle of Trump’s White House, “Fear,” Woodwa... 阅读全帖 |
|
l****z
发帖数: 29846 | 36 Democrats lost the South through culture war and elitism
By Timothy P. Carney | December 9, 2014 | 6:00 pm
If not for Southern Democrats, Republicans would have nearly had a
filibuster-proof Senate supermajority after the 2002 elections, giving
George W. Bush some real clout. Without Southern senators, Democrats wouldn
’t have taken over the Senate with Jim Jeffords’ party switch in 2001. If
not for Southern Democrats, Obamacare wouldn’t have become law.
For the foreseeable future, though, Democ... 阅读全帖 |
|
h*i
发帖数: 3446 | 37 From wikipedia:
"Y-chromosome haplogroup O3 is a common DNA marker in Han Chinese, as it
appeared in China in prehistoric times. It is found in more than 50% of
Chinese males, and ranging up to over 80% in certain regional subgroups of
the Han ethnicity.[28] However, the mitochondrial DNA of Han Chinese
increases in diversity as one looks from northern to southern China, which
suggests that some male migrants from northern China married with women from
local peoples after arriving in Guangdong, ... 阅读全帖 |
|
l*****a
发帖数: 38403 | 38 http://en.wikipedia.org/wiki/Christian_denomination
Catholicism
Main article: Catholicism
These are the churches which claim continuity (based upon Apostolic
Succession) with the early Church.
[edit]Catholic Church
Main article: Catholic Church
The Catholic Church is composed of 23 Rites: the Western or Latin Rite and
the 22 Eastern Rites (i.e., Eastern Catholic Churches).
[edit]The Latin Church
Main article: Latin Rite
The Latin Rite (not to be confused with the Roman Rite, which is one of the
... 阅读全帖 |
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m*********a
发帖数: 3299 | 39 Y-chromosome haplogroup O3 is a common DNA marker in Han Chinese, as it
appeared in China in prehistoric times. It is found in more than 50% of
Chinese males, and ranging up to over 80% in certain regional subgroups of
the Han ethnicity.[67] However, the mitochondrial DNA of Han Chinese
increases in diversity as one looks from northern to southern China, which
suggests that some male migrants from northern China married with women from
local peoples after arriving in Guangdong, Fujian, and other... 阅读全帖 |
|
p**p
发帖数: 10318 | 40 一堆堆的法律英文资料，看得我晕头转向，在youtube上顺便听音乐。。。一般的广告
我都不等开始就skip掉，这个忘掉了，没来得及。
一个南方口音，很像是Texas，田纳西那边的，一辈子在南方长大，甚至没离开过自己
的小town的老太太，说自己的一生，小时候在田里劳作，然后，怎么样，，，，，，嘟
嘟囔囔，还挺严肃的总结这一生，我主要听着南方口音很亲切，嗯，非常亲切，就没舍
得关掉，顺便听听她这一辈子家长里短。 结果老太太最后很认真的几句南方口音终于
把苦闷了一上午的我给逗笑了，大意，I lived long enough, and married three
times, yes, they all dead now, but I did not kill them, I just talked them
to death.
也祝大家新年快乐！！
再贴一首，我非常喜欢的Alan Jackson的Small Town Southern Man
"Small Town Southern Man"
Born the middle son of a farmer
And a small tow... 阅读全帖 |
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s**********t
发帖数: 1846 | 41 http://theeconomiccollapseblog.com/archives/what-is-the-best-pl
If you could live in any state in America, where would you go? During
troubled times like these, what is the best place in the United States to
live? A lot of people are asking these kinds of questions these days. Our
economy is on the verge of collapse, natural disasters are becoming more
frequent and more intense, the U.S. population is becoming angrier and more
frustrated by the day, our government has become incredibly oppressiv... 阅读全帖 |
|
b********n
发帖数: 38600 | 42 http://theeconomiccollapseblog.com/archives/what-is-the-best-pl
If you could live in any state in America, where would you go? During
troubled times like these, what is the best place in the United States to
live? A lot of people are asking these kinds of questions these days. Our
economy is on the verge of collapse, natural disasters are becoming more
frequent and more intense, the U.S. population is becoming angrier and more
frustrated by the day, our government has become incredibly oppres... 阅读全帖 |
|
p***y
发帖数: 18037 | 43 Taiwanese linguist races to save dying language
Photo: AP Children learn their tribal dialect in their aboriginal mountain
village of Piuma, southern Taiwan. In a race against time, a dedicated
Taiwanese linguist is trying to save the historically important aboriginal
dialect of Kanakanvu, spoken by only 10 people, from extinction.
Photo: AP Eighty-year-old Mu'u Ka'angena speaks to a graduate student,
documenting his dying tribal language in the aboriginal Kanakanvu mountain
village of Dakanua, ... 阅读全帖 |
|
I******a
发帖数: 3812 | 44 http://gyral.blackshell.com/names.html
Visit the original once in a while. Apparently, it's chock full o' updates.
Slur Represents Reasons/Origins
51st Stater Canadians Self-explanatory.
Abba-Dabba Arabs Used in the movie "Betrayed" by rural American hate group.
Abo Australian Aboriginal Derogative shortened term for Aboriginal.
Adolf Germans Hitler, of course. Generally thought of the worst person ever.
Ahab Whites From Moby-Dick. Ahab hunted the white whale.
Ainu Japanese Aboriginal Originall... 阅读全帖 |
|
w****r
发帖数: 245 | 45 大侠，分类都分好了，求神签
# Basic Materials, Agricultural Chemicals
AGU Agrium Inc.
AVD American Vanguard Corp.
CAGC China Agritech Inc.
CF CF Industries Holdings, Inc.
CGA China Green Agriculture, Inc.
CMP Compass Minerals International
COIN Converted Organics Inc.
IPI Intrepid Potash, Inc.
MON Monsanto Co.
MOS Mosaic Co.
NOEC New Oriental Energy & Chemical
POT Potas... 阅读全帖 |
|
B****2
发帖数: 8892 | 46 又找到一个网站，打着NCAA的名字，应该算比较权威的。
http://www.ncaa.com/history/football/fbs
按照其说法，ND有13个NC冠军，BAMA有12个。如此一来，如果BAMA获胜，两个队又平了
，有意思。
Championship History
YEAR CHAMPION SELECTING ORGANIZATION
2011 Alabama BCS
2010 Auburn BCS
2009 Alabama BCS
2008 Florida BCS
2007 Louisiana State BCS
2006 Florida BCS
2005 Texas BCS
2004 Southern California BCS
2003 Louisiana State, Southern California BCS, AP, FWAA
2002 Ohio State BCS
2001 Miami (Fla.) B... 阅读全帖 |
|
u***r
发帖数: 4825 | 47 http://www.nytimes.com/2011/12/27/business/global/chinas-push-f
GUANGZHOU, China — Three years ago, as part of its green-energy policy, the
Chinese government set an ambitious goal: by the end of 2011, the nation
would be able to produce at least 500,000 hybrid or all-electric cars and
buses a year.
With only about a week to go, it is clear China will fall far short of that
target. Despite dozens of electric-vehicle demonstration projects around the
country, analysts put China’s actual annual pr... 阅读全帖 |
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