f*******g
发帖数: 245 | 1 crisper/CAS9 究竟怎么工作？ homologous recombination ? or RNAi ? |
|
s*****t
发帖数: 107 | 2 各位大侠，小妹正在做Zhangfeng的LentiCrisper-V2 Vector，
已经按照ZhangFeng lab的Protocol把退火的Primer Pair连接到了
Lenti-Crisper-V2 Vector上。可是怎么知道我的Insert 连到vector上去了呢？
protocol说原载体上两个BsmB1之间有2Kb insert。 我现在把我连出来的Vector
作了个酶切，发现没有这个2kb insert了，但是怎么能排除原载体自连呢？
另外，我自己的insert连进去以后要不要考虑正反向的问题？ |
|
s*****t
发帖数: 107 | 3 Zhang Feng lab 给的Lenti-Crisper-v2 载体里面两个BsmB1位点之间有一个2kb
Insert。
是方便别人克隆用的，也就是只要把这个原来的２ｋｂ切掉，放入你自己退火的Ｐｒｉ
ｍｅｒＰａｉｒ就行了。我就是问连进去以后怎么鉴定？ |
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D*****a
发帖数: 810 | 4 CVS (1/1 -1/7):
Snickers Crisper Candy Bar (1.41 oz) - $0.99
Use $0.50/1 SNICKERS Crisper, Original, Almond or Xtreme Singles or Sharing
Size Bars (1.41 oz. – 3.59 oz.) printable
Pay $0.49, get $0.75 ECB for buying one (limit 1)
Submit receipt for $0.25/1 Snickers Crisper Candy Bar Ibotta rebate
Final Price: $0.51 money maker
Pampers Swaddlers Jumbo Pack Diapers - $8.99
Use $2/1 Pampers Swaddlers Jumbo Pack Diapers printable
Pay $6.99, submit receipt for $1.50/1 Pampe... 阅读全帖 |
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D*****a
发帖数: 810 | 5 Colgate Extra Clean Toothbrush (3 pk) - $3.99
Pay $3.99, get $3 ECB for buying one (limit 3)
Final Price: $0.99
Oral-B Indicator Contour Clean Toothbrush (2-pack) - $4.99
Pay $4.99, get $4 ECB for buying one (limit 2)
Final Price: $0.99
Buy (2) Oral-B Indicator Contour Clean Toothbrushes (2-pack)- $4.99
Pay $9.98, get $8 ECB
Final price: $0.99 each
(不知道能不能用 $2/2 Oral-B Adult Pro-Health or 3D White Manual Toothbrush
Packs，要是能用的话，会免费)
Irish Spring Body Wash (15-18 oz) - $2.99
... 阅读全帖 |
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D*****a
发帖数: 810 | 6 Clear Care Rinse & Go Solution 12 oz - $7.99
Use 25% Off Clear Care Rinse & Go Target Cartwheel
And use $5/1 Clear Care solution 12 oz+ printable
Final price: $0.99
Buy (3) Oral B 3D White Vivid Manual Toothbrush 2 pk - $4.74
Use 10% Off Oral-B Manual Toothbrush 2 count Target cartwheel
And use t(3) $2/1 Oral-B Adult Manual Toothbrush Multi Pack Printable
Pay $6.80, get $5 Target Gift Card
Final price: $0.60 per twin pack
Buy (3) Crest Pro-Health Toothpaste Twin Packs - $4.99
Use 10% Off Crest P... 阅读全帖 |
|
D*****a
发帖数: 810 | 7 CVS (1/8 -1/14):
Snickers Crisper Candy Bar (1.41 oz) - $0.88
Use $0.50/1 SNICKERS Crisper printable
Pay $0.38, submit receipt for $0.25/1 Snickers Crisper Candy Bar Ibotta
rebate
Final Price: $0.13
Buy (2) Cottonelle Bath Tissue 18 double rolls - $9.99
Use (2) $1.50/1 COTTONELLE Bath Tissue (18-rolls+) printable
Pay $16.98, get $5 ECB for buying two
submit receipt for $2 Cottonelle bath tissue Checkout51 rebate thru 1/11&#
8232;
Final price: $4.99 each
Buy (3) Brawny P... 阅读全帖 |
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o*****p
发帖数: 2977 | 8 这次的问题不是crispers, 是用crispers修饰生殖细胞。 |
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l****z
发帖数: 29846 | 9 by Jammie
Fresh off a four-year investigation into Sarah Palin’s uterus, Andy
Sullivan now has done has an in-depth look at his own colon. How’s the view
up there, Andy?
As the fall has turned crisper, a second term for Barack Obama has
gotten likelier. This may, of course, change: the debates, the Middle East,
the unemployment numbers could still blow up the race. At this point in 2004
, one recalls, George W. Bush was about to see a near eight-point lead
shrivel to a one-state nail-biter b... 阅读全帖 |
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D*****a
发帖数: 810 | 10 CVS (10/16-10/22):
Tide Pods (14 – 16ct) - $4.44
Use $2/1 Tide PODS 12 ct+ printable
Final price: $2.44
Buy (2) U by Kotex Barely There Thin Pantiliners (18 ct) - BOGO 50% off @$1.
77
Pay $2.65, submit receipt for $3/2 U by Kotex Ibotta rebate
Final Price: $0.35 money maker
Buy (2) U by Kotex Sleek Tampons (18 ct) - BOGO 50% off @$5.79
Use (2) $1/1 U by Kotex Sleek Tampons printable
And use (1) $2/1 U by Kotex Tampons CVS coupon
Pay $4.68, submit receipt for $3/2 U by Kotex ... 阅读全帖 |
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D*****a
发帖数: 810 | 11 CVS (10/23-10/29):
Buy (1) Tic Tac Mixers Cherry Cola (60 ct) - B2G1 free @$1.67
and buy (1) Tic Tac Fruit Adventure (60 ct) - B2G1 free @$1.67
and buy (1) Tic Tac Orange (60 ct) - B2G1 free @$1.67
and buy (1) Tic Tac Freshmints (60 ct) - B2G1 free @$1.67
and buy (2) Tic Tac Wintergreen (60 ct) - B2G1 free @$1.67
Use (3) Buy One Get One Free Tic Tac printable
Pay $1.57 submit receipt for $0.75/1 Tic Tac Cherry-Cola Ibotta rebate
and submit receipt f... 阅读全帖 |
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y*****o
发帖数: 689 | 12 产品信息直接抄过来。。。
The Emerson 4.6 cu. ft. (128L) compact refrigerator is perfect for small
offices or dorm rooms. Generous storage is provided with 3 interior racks, a
full width freezer compartment and door storage. The compressor cooling
system assures a consistently cool temperature with the environmentally
friendly R134a coolant which is also CFC free. Other features include an
interior light, separate crisper drawer, adjustable thermostat, reversible
door, flush back space saving design and manu... 阅读全帖 |
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h******y
发帖数: 351 | 13 用TALEN和ZFN的话off target的可能性也是有的。TALEN单侧靶位点一般设计16-18个碱
基，但是由于任何一个TALEN domain对相邻TALEN domain与靶碱基的结合是有影响的，
所以TALEN是可能结合到imperfect match的位点上去的。
由于TALEN的设计要在蛋白质水平进行，可以想象合成4^16种TALEN蛋白，从而对TALEN与
靶位点之间的特异性结合进行完全充分的研究是不可能的。所以实际上还没有文章像CR
ISPER那样对TALEN靶位点的每个碱基的重要性进行过系统的研究。相反，CRISPER只需要
合成oligo，单个碱基的突变很容易实现，所以能发现前面12-14个碱基最重要。但是这
并不等于说CRISPER只需要12-14个碱基，只是其余碱基的重要性相对较低而已。 |
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s******s
发帖数: 13035 | 14 非也非也。要看实际情况是啥，你这个是基础科研思路，只要knockout就
最好。实际上，很多疾病并不是knockout的，而是dosage造成的问题，有
可能更加接近RNAi。
应该可以1:1混合正常crisper：不切crisper造成hetero解决部分问题，
不过RNAi也可以装到inducible上做更加精细的表达调控，所以我觉得
很多情况下RNAi还是有特有的优势的 |
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t******s
发帖数: 935 | 15 俺记得gamespot对360的画面说不出啥优点，只好说360般的画面crisper。crisper是好
是坏就仁者见仁了。俺这回可能日英中三版都要入了，最后看看留哪一（几）个。 |
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V****n
发帖数: 651 | 16 王博士应该去接受一下Lubos Motl的教导 ：），或者Lubos的助教Xi Yin也可以。
文章没太大的问题，但不应该没有提到anthropic principle。我们居住的宇宙，正好
是10的500次方那么多个宇宙里面少数几个运气好，宇宙常数刚刚好可以产生生命的宇
宙的可能，也不完全是零.必须承认anthropic principle确实名声不好，不那么"
elegant".老爱好在死得早，否则一定会怀疑人生。
传统物理一向需要实验来验证理论，一直到相对论和量子力学建立的那几十年也是如此
。不过实验跟不上理论是迟早要发生的事情，正好我们赶上了。按我们现在的能力，你
总不能拿黑洞来做实验吧，就算做个地球直径一般大的加速器，离模拟宇宙开始普朗克
尺度也还差十几个数量级。这个时候是不是不能用实验来验证的理论就一定是玄学不是
科学，不是有正确可能的理论，那就难说得很了。
我倒是觉得也许我们应该等华大基因或其它机构研究发现到底哪些基因决定智商，然后
用crisper技术改造超级天才来发现新一代物理理论，可行的机会更大。毕竟目前研究
发现人类智商的上限在目前100之上30-100个标准差，... 阅读全帖 |
|
发帖数: 1 | 17 本来就不能组合，这种骗子是骗文科生的
能自由组合，千老那么费劲搞什么crisper |
|
发帖数: 1 | 18 不应该这么说。没有张，那个Doudna还在说真核能不能做不知道。是张发现了可以把
crisper用于真核细胞编辑 |
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a**i
发帖数: 608 | 19 10个类别里 TOYOTA 得五个冠军, HONDA 名落孙山
http://www.consumerreports.org/cro/consumer-reports-magazine/Ap
Top Picks 2012
The best models of the year in 10 categories
Consumer Reports Magazine: April 2012
A redesigned, significantly improved Camry Hybrid has helped Toyota capture
five of the 10 spots on this year’s Top Picks list. The other Toyotas that
made the cut are the Prius hybrid, the Highlander and RAV4 SUVs, and the
Sienna minivan. This is the first time since 2003 that one brand has so
dominated ... 阅读全帖 |
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h*****g
发帖数: 596 | 20 http://www.mitbbs.com/clubarticle_t/EasyMoney/31380171.html
Stores that allow buying third party gift cards with their own gift cards:
1. Barnes and Noble - many clerks will deny your attempt, but last time I
tried, the system did allow you to redeem Barnes and Noble gift cards for
some of the third party gift cards they sell.
2. Kmart - usually lets you use Sears/LandsEnd/Kmart GCs (they are all owned
by parent company, so you can use their GC's interchangeably) to buy third
party gift cards. E... 阅读全帖 |
|
s**********3
发帖数: 81 | 21 Whirlpool 双开门电冰箱 Refrigerator - 超级省电Energy Star Side-By-Side White
Excellent Condition
ENERGY STAR
Textured Door Design
Bow Handles
21.8 Cu. Ft. Total Capacity
Adaptive Defrost Control
EZ-Touch Crushed/Cubed Ice and Water Dispenser
Dispenser Lockout
Filtration System
Integrated Shelf Supports
Adjustable Spillguard Shelves
Clear Crisper
Clear Meat Pan
3 Adjustable Gallon Door Bins
Full-Width Freezer Bin
New sells for $1,000+ in store. We bought it new 6 years ago. This unit is
only lightly used a... 阅读全帖 |
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l********l
发帖数: 9452 | 22 head-fi review：
Details: Latest dynamic-driver offering from one of Head-Fi’s favourite
brands
Current Price: $149 from Head-Direct.com (MSRP: $149)
Specs: Driver: Dynamic | Imp: 150Ω | Sens: 95 dB | Freq: N/A | Cable: 4’ L
-plug
Nozzle Size: 5mm | Preferred tips: Large stock bi-flange, Meelec ‘balanced
’ bi-flange
Wear Style: Straight down or over-the-ear
Accessories (4.5/5) – Single-flange (3 sizes) and bi-flange (2 sizes)
silicone tips, replacement filters (5 pairs), and shirt clip; latest v... 阅读全帖 |
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w**********c
发帖数: 56 | 23 大家有遇到过相同问题的吗？怎么解决？看了一些网上评论，说法不一，没解决办法，
有的说要换压缩机，有的说把里面后面版卸下疏通。说这可能是samsung的通病。先谢
谢！ |
|
n***x
发帖数: 912 | 24 First and foremost, we would like to thank you for your continued excitement
and support for Plastc Card. We’ve had a tremendous response for our
product and we owe that, in large part, to our extremely supportive
community. It’s time for us to share a great deal of news - some exciting
and some that may be a little disappointing.
Plastc Card’s ship date is April 29, 2016. We realize that is a
considerable delay from our initial projection. We apologize for making you
wait and assure you that we... 阅读全帖 |
|
y**y
发帖数: 1261 | 25 原贴2010发的，也许不够update, 有些可能已经不适用了, 仅作参考，欢迎大家踊跃更
新，
小白鼠积极出洞，没有Safeway 也有省版Pennyway.
Barnes and Noble 这个我试过不行。
_____________________________________________________________________________
可用
Stores that allow buying third party gift cards with their own gift cards:
1. Barnes and Noble - many clerks will deny your attempt, but last time I
tried, the system did allow you to redeem Barnes and Noble gift cards for
some of the third party gift cards they sell.
2. Kmart - usually lets you use Sears/Lands... 阅读全帖 |
|
n********5
发帖数: 513 | 26 Stores that allow buying third-party gift cards with their own gift cards (
in-store purchase) Updated 24 November 2014 - Please put verification next
to updated line with date:
1. Barnes and Noble - Most clerks will deny your attempt, supposedly the
system will allow you to redeem Barnes and Noble gift cards for some of the
third-party gift cards they sell.
2. Sears - You can buy Sears/Kmart gift cards and pay with Sears/Kmart gift
cards. They have many including Shell, Staples, Subway, and mor... 阅读全帖 |
|
D*****a
发帖数: 810 | 27 Target coupon: buy $15 Skin Care Purchase (Facial Skin Care, Natural Skin
Care, Hand & Body Lotion), get $5 Target Gift Card (in-ad Target coupon or
text GLOW to 827438)
Buy (2) Differin Acne Treatment Gel 15 g - $12.99
Use Free $5 Target Gift Card with $15 Skin Care Purchase Target Coupon
Pay $25.98, get $20 Gift Card back for buying 2 thru 1/14
and get $5 Target Gift Card for $15 Skin Care Purchase
Final price: $0.49 each
Buy (4) Aquaphor Healing Ointment Tube (1.75 oz) - $4.99
Use Free... 阅读全帖 |
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a****d
发帖数: 1919 | 28 赞专业分析
补充一点，JUNO几年前和EDIT签了合作协议，利用EDIT的crisper-cas9技术对CAR-T制
品进行基因编辑，不过具体进展一直没看到消息。
CRS
CD19
别。 |
|
Y**i
发帖数: 3016 | 29 Altec Lansing GT5051 Computer Speaker 5.1 - $25 obo
in excellent condition
$25 (MSRP: $150)
please email me at PurdueUSA .at. gmail.com or call me 765-409-8119 if
interested
see more info and reviews here:
http://www.bing.com/shopping/Altec-Lansing-GT5051-PC-multimedia-home-
theater-speaker-system/search?q=altec%20lansing%20gt5051&p1=
[CommerceService+scenario%3d%22s%22+docid%3d%22B391DE8CEB478D5874A1%22]&
wf=Commerce&qpvt=altec+lansing+gt5051
SPECIFICATIONS
The GT5051 is a genuine 5.1 surround ... 阅读全帖 |
|
n***s
发帖数: 10056 | 30 "Canola / rapeseed oil
One little-known source of trans fat is canola / rapeseed oil. The trans fat
occurs as a result of processing, which takes place at high temperature.
The raw seed begins with a high level of beneficial omega-3 oils, however
these tend to oxidise during processing producing off, rancid odours. During
deodorisation, some of the omega-3 fatty acids are converted to trans.
The proportion converted to trans is highly variable - in general, UK oils
have low levels of trans, howe... 阅读全帖 |
|
w******0
发帖数: 4472 | 31 多谢！ 长见识了！
"Canola / rapeseed oil
One little-known source of trans fat is canola / rapeseed oil. The trans fat
occurs as a result of processing, which takes place at high temperature.
The raw seed begins with a high level of beneficial omega-3 oils, however
these tend to oxidise during processing producing off, rancid odours. During
deodorisation, some of the omega-3 fatty acids are converted to trans.
The proportion converted to trans is highly variable - in general, UK oils
have low levels of ... 阅读全帖 |
|
s*********e
发帖数: 90 | 32 I like XD better:
1)I like the addition of grip and drop safety.
2)glock trigger and sights are plastic, XD are steel. XD has crisper
trigger break |
|
|
G********A
发帖数: 2851 | 34 Amaryllis Planting and Care
After-Bloom Care
After the amaryllis has stopped flowering, it can be made to flower again.
Cut the old flowers from the stem after flowering, and when the stem starts
to sag, cut it back to the top of the bulb.
Leaf Growth and Development. Continue to water and fertilize as normal all
summer, or for at least 5-6 months, allowing the leaves to fully develop and
grow. When the leaves begin to yellow, which normally occurs in the early
fall, cut the leaves back to abou... 阅读全帖 |
|
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a***n
发帖数: 1500 | 36 第一张是索蔡合影。
能看到酒红色T*镀膜的是索蔡2470/85/135。这几只镜子也是使用频率最高的。
Zeiss镜头盖底下的是小弟16-80/3.5-4.5 DT，已经和A700一起出掉了。
这张是最丧心病狂时期的全家福，三只索蔡也在其中：
这张是135/1.8出的St Patrick's Day Tap Dance
*****
懒人一头，从FM抄了一段T*历史过来
Taken from http://www.fredmiranda.com/forum/topic/215494
Starting with a T* coating definition
Carl Zeiss T* anti-reflection coatings ensure the virtual elimination of
unwanted reflections inside the lens. This means that only image-forming
light reaches the image plane of the camera. Your photographs will have
g... 阅读全帖 |
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H********g
发帖数: 43926 | 37 再一个问题是很多paper根本没重复的价值。就一个耸人听闻的概念，对其他人的研究
并没有什么实质影响。而且有的概念还方便别人写review，就算是假的也没人有动力去
揭发。
小宝这个东西是因为使用价值太高所以被一堆人重复了。造假的大忌就是去编造一种可
以当工具的东西。生物里被人检验了无数次的东西还是有的，比如限制性内切酶，PCR
，去年的crisper。 |
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H********g
发帖数: 43926 | 38 再一个问题是很多paper根本没重复的价值。就一个耸人听闻的概念，对其他人的研究
并没有什么实质影响。而且有的概念还方便别人写review，就算是假的也没人有动力去
揭发。
小宝这个东西是因为使用价值太高所以被一堆人重复了。造假的大忌就是去编造一种可
以当工具的东西。生物里被人检验了无数次的东西还是有的，比如限制性内切酶，PCR
，去年的crisper。 |
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i******n
发帖数: 538 | 39 A long article from New York Times in memory of Sutherland's life and career.
Joan Sutherland, Opera Soprano, Dies
By ANTHONY TOMMASINI
Joan Sutherland, one of the most acclaimed sopranos of the 20th century,
died on Sunday at her home in Switzerland, near Montreux. She was 83.
Her death was confirmed by her close friend the mezzo-soprano Marilyn Horne,
a spokesman for the Metropolitan Opera said.
After the Australian-born Ms. Sutherland’s Italian debut in Venice, in 1960
, the country’s notorio... 阅读全帖 |
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w********1
发帖数: 3492 | 40 Tue, 12 Jun 2012 10:45:13 PDT
AnandTech takes a closer look at the new MacBook Pro's 2880x1800 Retina
display, revealing just how well the display stacks up against its
predecessor and other notebook displays and delving into the details of how
it handles various resolutions.
In looking at scaling, the report explains the new slider option in System
Preferences that allows users to select from a spectrum of resolutions that
include not only the 1440x900 resolution in Retina quality using the ful... 阅读全帖 |
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C***d
发帖数: 3779 | 41 The beauty of having lots of Windows 8 review units lying around is that it'
s easy for us to compare all of those touchscreens side by side. Even amid
so much competition, the Yoga 13 comes out looking pretty good. The 1,600 x
900, 300-nit IPS panel looks noticeably crisper next to more pedestrian 1,
366 x 768 displays, though it's still not as stunning as the 1080p panels
used on the XPS 12 and Acer Aspire S7. Then again, those two machines each
have a higher starting price, so is anyone reall... 阅读全帖 |
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H*******i
发帖数: 196 | 42 原核里面off-target 比例也类似么？ 弄一堆容易出现的off-target序列，放在生存
必须的基因里，但是target的序列条件致死（比如和一个筛选因子结合后致死）。然后
error-prone PCR crisper系统 电转不知
道可以不。。 |
|
j********r
发帖数: 156 | 43 扫了一下elife的文章，感觉像molecular版本的current biology。尽管她不care IF,
第一次能上10就是miracle了。再看看cell 2011 Weinberg的the hallmarks of cancer
,citation都上5k。
nature那些TCGA的文章都around 0.2k。elife这样的star story太少了。年初的那篇
crisper刚50。
(当然我们的story如果elife能看上，也是一个miracle)
btw, take a look at this paper
http://elife.elifesciences.org/content/2/e00542
what a miracle, again! |
|
m******g
发帖数: 467 | 44 Part8 感觉这个帖子就是抒发感想用的吗……
首先说明一下，楼主是长期遭受日漫荼毒的中二青年，所以想法和说法都会非常中二。
最近国内有小朋友想申请出国，所以来询问我，我也就将所知情况一一告知，并未加上
任何抒情体。末了，他有点不好意思地追问“在国外读博开心吗？”，我说“挺好的啊
，至少我95%的时间没有后悔出来，如果不出国，估计我30%的时间会臆想要是出国就好
了”，他追问“可是我看了网上好多人对生物博士/后都黑得很厉害呢”，我回答“真
正的牛人是不会在网上炫耀说自己过得有多幸福，因为说了，肯定会有人跟帖说‘再等
几年你就知道厉害了’，所以他们不会自讨没趣的，而那些说自己过得不爽的往往能引
起其他人的共鸣，有一些心理安慰的作用呢”，他觉得也有点道理，现在还在申请中，
未知结果如何，个人觉得他有自己的想法，有很勤奋，学生物是很适合的。
写这一Part的目的就是奉劝生物黑不要去吓小朋友。“21世纪是生物的世纪”这句话现
在已经被很多人当做笑话来说了，其实这句话基本没错。去看看每周的CNS，PNAS等等
，你就知道生物是多么的日新月异，稍微了解一下各种测序技术、CRISPER等等，你就
会... 阅读全帖 |
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n********k
发帖数: 2818 | 45 这应该同CRISPER没得比！筛选达到100%，不双筛呢！而且同源臂长很多了，而且与具
体位点影响大了。我的KO单选，ES里面基本效率是20-50%, 为什么，ES 高表达呀。同
理在这篇文章了，所以复杂着呢。 |
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s******s
发帖数: 13035 | 46 两回事。crisper可以做homozygous mutant，也就是knockout；
RNAi这玩意儿更像hetreozygous mutation，knockdown |
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j****x
发帖数: 1704 | 47 尽管是两回事，但假如crisper做knockout能和RNAi做knockdown一样容易，那么的确可
以预计前者将会取代后者吧 |
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r******k
发帖数: 446 | 48 如题。 小弟想想做cell line的KI。不知道哪位大神做过？ 质粒我都拿到了。 不知道
大家都用什么样的protocol呢？
非常感谢 |
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