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全部话题 - 话题: cloning
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l**x
发帖数: 296
1
来自主题: SWJTU版 - clone and stone
A mad scientist named Stone became obsessed with cloning. He tried to clone
everything he could get his
hands on. When he ran out of cloning subjects, he decided to clone himself.
He finally achieved what other
scientists thought was impossible; he cloned himself, and he was beside
himself. This led to serious problems.
Stone and the clone didn't get along because they were both insane, another
name for mad. They began to
argue about who was Stone and who was the clone. Stone decided to
w*******y
发帖数: 60932
2
Stopped by my local Target and found an awesome mark down on the Lego
StarWars Clone Turbo Tank. Reg $119.99 marked down to $29.98!! Picked up
both they had sitting in clearance.
Its this one - Link:
http://www.target.com/p/Lego-Star-Wars-Clone-Turbo-Tank/-/A-123
#?ref=tgt_adv_XSG10001&AFID=Froogle_df&LNM=%7C12338187&CPNG=toys&ci_src=
14110944&ci_sku=12338187" rel="nofollow" target="_blank" onclick="_gaq.push(
['_trackEvent', 'thread', 'click', '3334284 - target-clearance-lego-starwars
-clone-... 阅读全帖
g****y
发帖数: 323
3
来自主题: _America版 - 30,000秘密的继续(Clone)
昨天看PBS Rose的访谈节目,关于human clone.
里面一个也提到了30,000, 因此我也就姑且认为这个数据正确。
节目中有两个教授,一个记者,一个活动家(I forget what he is
doing, but not a researcher). 两位教授,和活动家都表示不宜现
在搞human clone, 记者则极力鼓吹human clone, 表示从他得到的消
息,这件事的危险是可以控制的,这件事的意义是伟大的。当然
他除了不点名的说有人这么告诉他,同时又强调对于不孕夫妇的伟大
意义,在技术上他是一点也说不上来。两位教授到是谈了一些,包括

1.human clone,在极短的时间内完成胚胎发育可能几个月后才会有
的功能(指某种蛋白的出现?),这样的reprogramming的过程是违
背自然的胚胎发育的,可能有无法预测的缺陷。
2.并非所有的基因都得到表达,有些基因可能日后才会表达,更增加
可能存在缺陷的可能。
3.动物clone的时间不长,成功率不高,sheep, mice是有一定的成功
率,但是在猫狗身上还从来没有。即使是表面的行为,有些clone
a
w********p
发帖数: 948
4
不好意思,很丢脸的来问一句。
9.10 you have a stack of n boxes, with widths w, height h and depth d. The
boxes can not be rotated and can only be stacked on top of one another if
each box in the stack is strictly larger than the box above it in width,
height and depth. Implement a method to build the tallest stack possible.
where the height of a stack is the sum of the heights of each box
career cup 9.10堆盒子。答案的最后一句为啥是return (ArrayList)max_stack.
clone();
一般来说,只有在会 expose internal reference object 才会用retur... 阅读全帖
z****e
发帖数: 54598
5
因为现在都讲究singleton等控制,你用clone是啥意思呢?
clone本质上就是一个new,而且是一个无法控制的new
比如你private构造器可以不让一个对象随随便便new出来
但是private构造器无法控制clone方法
这个其实是一个不受控制的使用,clone是一个pattern
不是说避免使用,而是用得很少而已,一般都是写框架的人不推荐你用
因为框架无法控制,框架经常要管理beans的生命周期
从诞生到消亡,这个它控制不了,所以就不推荐你用
b******s
发帖数: 1089
6
来自主题: Biology版 - 再问gateway cloning
本来容易的gateway cloning成了最近几个月摧毁我身心健康的罪魁祸首。
哎,说来话长。先来介绍背景:1年前开始用gateway做克隆。在此之前实验室也一直用
gateway,但是实验室很小,我之前只有一个日本博后。做了好几年。几乎所有的东西
都是从他那里来。我们用的是multisite gateway,细说就是三个entry clone和载体一
起连。
我做了N次(我都不好意思说N等于多少了),一个确定的都没成功。
症状:每次连接之后转e coli都有菌落,而且菌落看起来正常。介入液体摇也正常生长
(spectinomycin resistance)。但是colony PCR有时会有不浓的bands,size貌似正
确,但是很多时候PCR失败。提plasmid PCR也失败,酶切也失败。
可能一:entry clone有问题。用gateway做entry clone很容易,应该不会,我做了好
几个entry clone,都sequence过,PCR检测等很正常。
可能二:我哪里做错了。至少我现在想起来应该没有。我经过无数次改进,从反应DNA
浓度到反应时间到heat shoc... 阅读全帖
r******t
发帖数: 2
7
来自主题: Hardware版 - Disk clone 问题
Laptop 的硬盘出了点问题需要换一个。以前都是按步就班,重装系统,drivers, 软件
, 拷贝数据...
现在硬盘上东西实在太多了,按步骤来一遍太花时间,所以考虑Disk Clone。以前从来
没有Clone过,想问一下Clone有没有什么潜在的问题。Clone时有什么需要注意的吗?
谢谢!
U*****n
发帖数: 33
8
来自主题: Hardware版 - Clone SSD with bad sectors?
FangQ,
I tried ddrescue (a Linux utility) and it cloned the defective SSD and
claimed a success. ddrescue reported errors of 2MB. But somehow when I tried
to reboot the computer it did not boot from the clone (I did choose the
clone as the boot device using the F12 key).
I tried Clonezilla that runs on a Ubuntu live USB, but it stopped working
due to disk errors.
Have you had any experience with the above utilities?
Let me make sure I understand your suggestion:
1. Make a Ubuntu live USB
2. inst... 阅读全帖
c**r
发帖数: 211
9
来自主题: Hardware版 - clone软件求推荐
一个1TB的系统盘,要clone到一个250GB的 SSD
系统盘只用了200GB左右,但免费的clone 软件比如clonezilla不能大clone到小硬盘。
求推荐个免费的可以大硬盘clone到小硬盘的软件,多谢!
p***t
发帖数: 698
10
Thinking in Java说Object的clone是比较特殊的。意思是违反了类
继承的原则。
clone在Object中是protected的。比如编一个如下的程序:
Public Class B {
public static void main(String[] args) {
A aa = new A();
aa.clone();
}
}
Class A {
}
Suppose应该是A继承Object中这个protected的clone的。这种特殊是
啥个机制呢?
c******n
发帖数: 4965
11
来自主题: Java版 - git clones only one branch?
if I do
git clone git://git.apache.org/cassandra.git
it gives me only the trunk branch, but they have many branches
I have to use the -b arg for clone to get the other branches.
"clone" clones a "repository", doesn't "repository" mean the collection of
all branches?
now, since I can't have all branches in my local repo, it's difficult to
compare the differences between trunk and another branch
r**h
发帖数: 6
12
请教有关专家:
在Sanger没有找到该位点的129的BAC clone,还有什么library可以推荐?比如说RPCI-
22怎么样?不过我没找到定位single clone的bioinformatics工具,比如ensembl之类。
不准备自己做hybridization to screen the clone,因为花费力气和金钱比较多;
也不希望直接打C2 ES cell line,担心效率会有所下降;
也不知道在该位点,129和C57的区别会有多大,不知道用C57的BAC做的construct打129
ES cell,会不会比较不好;
所以回到开始的问题:如果执着于129 BAC clone,除了Sanger,还有什么library选择
(如果不自己筛的话)?或者说,应该执着于129 BAC吗?
完全新手一个,简单问题比较多,贻笑大方了。谢谢!
l****n
发帖数: 3
13
来自主题: Biology版 - stable clones vs. stable pool
Then I have another question. What are the best controls for the stable
clones? Theoretically, it would be the stable clones with the empty vector.
But cancer cell lines are heterogeneous at the first place. After
antibiotics selection, I might end up having control clones with different
phenotypes. Which control clone(s) should I then use?
q**********0
发帖数: 335
14
Recently I tried three different TA cloning Kits from Invitrogen (TA cloning
, TOPO TA clong and dual promoter TA cloning). I found all of them have very
high self-ligation rate (>90%). I don't know anybody have similar results?
Any suggestions for other better choice for TA clone? Thanks.
w*******y
发帖数: 60932
15
Amazon
Star Wars Clone Wars Electronic Blaster - Clone Trooper
= $9.99
Features a targeting light
The epic fight is heating up
Grab your blaster so you?re ready
Thrilling blaster modeled after the Clone Trooper characters? blasters
features a targeting light and blasts foam suction cup-tipped darts
Blaster comes with 3foam darts
K**i
发帖数: 333
16
来自主题: EmergingNetworking版 - clone mac address 遇到问题 (无法save)
用的是BELKIN的,上面有MAC ADDRESS, 然后有 clone的功能。当我clone的时候,MAC
ADDRESS空格的显示的是PC的MAC,是对的,没问题。但是如果我重新再查看MAC ADDRESS
的时候,又变回原来的MAC了,就是说不能KEEP我CLONE后的MAC ADDRESS. 是怎么回事
呢?感觉很是STUPID。总觉得哪儿有问题,却不知道到底哪儿出了问题。
还有一个问题就是,用这个ROUTER直接用网线联电脑的时候(还没敢奢望无线),每次
大概能联10秒钟的样子,ROUTER显示CONNECTED.然后网络就不通了,ROUTER上也显示NO
CONNECTION了。是怎么回事呢?
k*****y
发帖数: 744
17
来自主题: Hardware版 - 问一下dd-wrt下mac clone的问题
之前没开mac clone,先开modem,过一会之后再开router是能得到wan ip。不过好像经
常renew lease的时候就会丢了wan ip,然后再也连不上了。
昨晚开了mac clone,clone了x61t的mac,现在好像没有这个问题了。请问这是什么原
理?ISP对电脑的mac宽容一点?
t**r
发帖数: 3428
18
java里是不是要避免用clone()有人说clone() is broken.
看看大牛的看法?
b******e
发帖数: 61
19
最近被一个cloning给弄得焦头烂额的。我的目的是想把一个4kb的片断从pcDNA3弄出来
,然后克隆到pLenti7.3里边去。我试了两种strategies:
1, 先把4kb的片断从pcDNA3用RE切出来,然后用klenow补平,再用Taq加上3’A。然后
用TOPO TA cloning到pLenti7.3。结果:一个克隆都没有长
2, 用PCR直接把4kb的片断从pcDNA3里amplify出来 (用invitrogen pfx DNA
polymerase),然后用Taq加上3’A,再用TOPO TA cloning到pLenti7.3。结果:长了一
个克隆,假阳性的。
想请教各位大牛,有没有什么建议对于大片段的TA连接。万分感谢!!!
X***n
发帖数: 366
20
来自主题: Biology版 - Difficult cloning - 求助
有BAC clone 吗?有的话基于BAC clone recombineering delete掉不需要的部分就行
了。
PCR clone是够呛的,基因组不大的话自己做BAC未必不是个选择。
S**********e
发帖数: 1789
21
来自主题: Biology版 - stable clones vs. stable pool
这个很难讲。stable clone 如果只有1个,那你看到的有可能是插入了某个重要区域后
的表型。相反的结果也有可能。stable clone 是真的,你的pool里面可能这个稳定的
外源基因占少数,被掩盖了。估计你要多弄几个stable clone, 然后比较一下。

stable
n********k
发帖数: 2818
22
make sure it is what you said, if yes, may consider to change bacteria
strains, look for those which can overexpression long repeats DNA, or
inverted repeats, or for retro and lenti viral stuff...essentially those
lost RecABCD, not the only the RecA as the typical strains..if it is true,
this kind of cloning work is not trivial until you figure out right
materials...Or you could beat the nature by brutal force, I heard a guy
picked 20K clones to get one construct/plasmid done, that was before we... 阅读全帖
p***e
发帖数: 35
23
弯区大制药厂,想找个擅长molecular cloning的,专门做clone. 唯一要求就是喜欢而
且安心给整个department 的人做clone. 技术新,做的快。
w*******y
发帖数: 60932
24
Link:
http://www.bdlot.com/dvd-clone/giveaway.htm
more info on the product- Link:
http://www.bdlot.com/dvd-clone/


quote



Create Your Own Perfect DVD Backups of Raw Content with Ease
Remove All Known DVD Protections
Clone Entire DVD to Blank DVD
Backup Full Content of DVD to Hard Drive or USB
Backup DVD Main Movie to Save More Space
Burn to Blank DVD in an Easy and Fast Way
Flexible Virtual DVD Drive Simplifies Your Work
Supported DVD Copy Protection:
1. Reg... 阅读全帖
m**i
发帖数: 724
25
来自主题: _FreeSpace版 - Star Wars: Attack of the Clones
Star Wars: Attack of the Clones
HOST:
Movie fans are enjoying the new Star Wars movie, "Attack of the Clones." It
has earned more than two-hundred-fifty-million dollars since its release five
weeks ago.
[Image Removed]
The movie is the fifth in a series written and directed by George Lucas. They
tell a continuing story about a young man named Anakin Skywalker in an
imaginary galaxy of planets. Shirley Griffith has more.
ANNCR:
In "Attack of the Clones," a group of separatists wants independence
A*******e
发帖数: 12486
26
来自主题: Military版 - China Clones, Sells Russian Fighter Jets
ZHUHAI, China—A year after the collapse of the Soviet Union, a cash-strappe
d Kremlin began selling China a chunk of its vast military arsenal, includin
g the pride of the Russian air force, the Sukhoi-27 fighter jet.
For the next 15 years, Russia was China's biggest arms supplier, providing $
20 billion to $30 billion of fighters, destroyers, submarines, tanks and mis
siles. It even sold Beijing a license to make the Su-27 fighter jet—with im
ported Russian parts.
Today, Russia's military bonan... 阅读全帖
d********w
发帖数: 363
27
来自主题: JobHunting版 - 一道fb的题,clone a graph
大家看我这样写对么,dfs来做,map来保存是否之前遇过。
1 struct Node {
2 int val;
3 vector neighbors;
4 }
5
6 Node* clone(Node *root, HashMap& map) {
7 if (root == NULL) {
8 return NULL;
9 }
10 if (map.find(root) != map::end()) {
11 return map[root];
12 }
13
14 Node* p = new Node();
15 p->val = root->val;
16 p->neighbors = new vector();
17 vector& v = p->neighbors;
18 for (int i=0; ineighbors.size(); i++... 阅读全帖
W**********r
发帖数: 8927
28
来自主题: StartUp版 - China's Facebook Clones
http://www.readwriteweb.com/archives/china_facebook_clones.php
China's Facebook Clones
Written by Gang Lu / June 24, 2008 11:00 PM / 7 Comments
Facebook has launched a Japanese version and a Chinese version (the latter
announced this week). However, many Facebook clones have been in operation
in China for a long time. So whether Facebook has an official presence in
China does not really matter for millions of Chinese users. Perhaps you are
bored with these China copycat stories. However if we st
s******3
发帖数: 1767
29
我是把新硬盘先放进电脑,旧硬盘装到硬盘盒,然后从光盘启动clone软件 (Acronis
True Image
bootable disk),clone硬盘的
b******n
发帖数: 823
30
来自主题: TVGame版 - xbox clone法换大硬盘
之前follow老鳄鱼的换大硬盘攻略,其他功能都没有问题,
只要一进入正版游戏,马上报错误21。
无法,只能做他想。晚上折腾了几个小时,终于搞定。
将已经softmod过的硬盘和要换的大硬盘都unlock,然后接在IDE上,
(需两根IDE线,光驱还要占一个口)
进入xboxhd之后,第三项是Clone existing Xbox HD,选择好sourse和destination,
就开始dd了。
这段过程比较长,我10G硬盘花了1个多小时。
完成之后把大硬盘重新lock,dah dah。 done。
这个方法好处是简单,softmod后不需要做第二张光盘,缺点是慢,需要机器有俩IDE口。
另外,搞定后google了一下,有人提到可以用ghost bit by bit的clone,我相信比dd
快,
有兴趣的也可以去试试。
b********t
发帖数: 992
31
'Clones' DVD May Hit Stores Quickly
Wed May 15, 8:31 AM ET
By The Associated Press
"Star Wars: Episode II — Attack of the Clones" will be available on DVD this
fall, the franchise's creator, George Lucas, said last week in an interview at
his film headquarters, Skywalker Ranch.
"Episode I — The Phantom Menace" came out on videotape early in 2000, about
10 months after the film's theatrical debut, but the DVD version was not
released until a year and a half later. The delay resulted because Lucas
y*b
发帖数: 3190
32
zz
The Yongnuo YN 35mm f/2 clone lens for Canon DSLR cameras is now in stock at
Amazon ($122) and eBay ($116). Here are the lens technical details:
The closest focusing distance up to 0.25 meter,when under close-up shooting,
it is available to get the unique sense of depth of the wide-angle lens.
F2 large aperture,virtual background,which makes photography with more fun.
Support auto focus (AF) and manual focus (MF) mode.
The aperture blades up to 7 pieces,which allows you able to shoot the near... 阅读全帖
s*****d
发帖数: 160
33
来自主题: XJTU版 - 想起了clone这个问题
据说现在中国正在研究用CLONE的方法保护大熊猫,
我不知道这个方法是否真的管用,不过据我看来,
外国人的很多方法都只治表不治本。假如CLONE只是
完全复制了母体的DNA组合序列,物种不变化,就
违反了自然规律,又如何能发展生存呢?就象人类
如果一直停留在农业社会,手工业社会,不变化,不
发展就不会有延续的可能性。
由此想到发达国家搞了许多发明创造,蒸汽机使工业
革命到来,信息技术使产业革命到来,但是这些技术也
产生了诸多问题,我在以前的文章中已经叙述过。
我们是有几千年历史的文明古国,不是我自夸,有历史
的国家做事情毕竟成熟一些,国人性格上缺少冲动,冒险
的精神,也好也不好。虽不象美国人那么爱竞争,爱
挑战,喜欢变化多端,但是步子走得似乎稳一些。要
研究历史有许多值得深入的地方,但可以通过一些实际
的例子来进行对比:
比如西医与中医。对付外伤,比如伤残,肿瘤等等,西医见效
来得个快,但是对付慢性疾病,中医以其自然稳妥的疗法更胜
一筹。西药的付作用更大,好比工业革命带来的付作用也是
更大的。西医在考虑某种疾病的时候往往头痛医头,脚痛医脚,
不象中医,头痛的时候想到肾是不是出了毛病,胃
l*******d
发帖数: 93
34
来自主题: CS版 - how to clone dell os?
how to clone dell os?
I bought a DELL XPS. There is no Windows CD included. How to clone the current
operating system only rather than the whole big hard drive? So that I can
restore the system in case of crashes. Thank you!
y**b
发帖数: 10166
35
以前旧电脑一直都是用clone模式,通过s-video将电脑里面视频
送到TV,电脑干活不受影响。
现在一台dell vostro 410/HD3450/24"LCD 1920x1200,通过hdmi
连接到42" 720p/1080i LCD TV, 也是clone mode,结果只要hdmi
一插上,就把24"LCD(无论dvi还是vga连接)的分辨率降到1920x1080,
只有拔掉了才能恢复1920x1200. ATI Catalyst里面找不到独立
控制分辨率的选项。
简单说,我希望24"LCD里面始终保持1920x1200,而TV那边分辨率
不是很在意(因为大部分视频都不是高清)。不知道有没有办法?
另外一插上hdmi就降到1920x1080是不是跟电视机有关?
h**2
发帖数: 2841
36
clone 还是 extended display? clone应该是两个完全一样的吧
试试用extended display?这样电视机应该可以用不同分辨率
eg. 笔记本是1280*800,电视机是1024*768,不过是svideo的
y**b
发帖数: 10166
37
我用clone,以前lcd只有1680x1050,输出到tv没问题,
现在lcd 1920x1200再想输出到tv就出现这个问题。
extend模式可以工作,但是远没有clone模式方便(任何时候只要打开视频,
电视立即播放,不必拖拽、最大化,而且lcd里面也可以同屏随时观看)
b*******r
发帖数: 6655
38
汇报一下使用经历
根据评测先后装了
Macrium Reflect
Paragon
安装都很简单,就是paragon需要注册(free)
前者30M, 后者100多,据说可以做differential backup and partitioning
装完以后发现不能直接clone, 必须先作img,而且这个paragon 仅仅从65G压缩到50G,
就显示需要2.5 小时
最后不得已到seagate 下了disc wizard,就是Acronis True Image 的简化版,使用非常简单,直接clone,就是没法自由分区,
只能按源盘的比例来, 可以缩小,剩下空间的重新Format为另一个分区,就是说你原
来两个区,新的必须四个区。非常可笑。好处是比较快,65G总共半小时搞定。
a*****n
发帖数: 14370
39
来自主题: Hardware版 - clone多台电脑 SSD
请教一下大牛们。如果我有50台desktop或者laptop,想把全部普通硬盘升级成 SSD,
有什么简单一点的办法?最直接就是一台一台来clone,不过这也太痛苦了。
因为这批电脑所有配置完全是一模一样的,可不可以把其中一个HDD拆出来,
把这个HDD clone到50个SSD上面。然后再把SSD接到各台电脑上面。
问题来了,这所有50个SSD里面的东西一模一样,windows 10 product key也是一样,
这样可以吗?不过听说win10 key是存在主板上面的,或者这样也是可行的?
如果上面这个方法可行,那我就是用一个external HDD doocking 和一个 SSD USB
transfer kit来完成。因为是usb,应该也是相当慢的了。或者有各好的方法?
请大家指点迷津,大包子感谢!
F***Q
发帖数: 6599
40
来自主题: Hardware版 - clone软件求推荐

first, use a partition editor to resize the 1TB drive down to 250GB (just
slightly less than the target disk), then, clone.
I have always used gparted+dd for my disk clones, never had any issue.
https://www.tigraine.at/2014/01/04/how-to-upgrade-to-an-ssd-with-minimal-
effort
m******t
发帖数: 2416
41
来自主题: Java版 - Why Java needs clone interface?
IMHO, the actual reason is Java has to let the programmer decide
whether to make deep or shallow copy while cloning. In some cases,
a hybrid of deep and shallow copy is also possible.
Also, while it's not hard for Java to provide a generic clone utility,
it would have to be based on reflection (similar to how serialization
works), which could be faily expensive operations.
l**********n
发帖数: 8443
42
干嘛要clone,serialize,再deserialize
g*****g
发帖数: 34805
43
Nothing wrong with clone, but you have to mark the interface and remember it
's shallow copy by default.
f*******n
发帖数: 12623
44
凡是说X是broken的人都是不懂X怎么用,或以为X是来做Y的,发现做不了Y,就说X
broken。你如果真正明白clone是什么你就可以自己决定符不符合你的要求。
documentation没有骗你。
l*******d
发帖数: 93
45
来自主题: Windows版 - how to clone dell os?
how to clone dell os?
I bought a DELL XPS. There is no Windows CD included. How to clone the current
operating system only rather than the whole big hard drive? So that I can
restore the system in case of crashes. Thank you!
g*******t
发帖数: 1039
46
来自主题: Windows版 - OS disk clone
I have an old drive, partitioned into C: and D:
It's old so I got a new drive and trying to clone the old drive using Ghost9.0
I completed clone old C:\ (Ghost shows the task completed), and the old drive
died.
I swapped the new drive in, it seems working, but as soon as I typed in
username/password, it automatically logs off
any suggestion?
thanks!
l*****k
发帖数: 587
47
来自主题: Biology版 - another clone trick: 3-way ligation
Well, I am sure I am not the first to do this, but I do want to share it
Sometimes, cloning multiple things into a vector can be tedious as you
have to transfer the fragments through several vector to get the
desired one, usually because the existence of the same Restriction site
in the vector and the fragment you want to clone.
say a gene has 2 frags in two separate vector, now you wanna get
them ligated into the same vector.
EcoRI------------XhoI Frag I (has interal BamHI site)
XhoI-
g***y
发帖数: 201
48
多年没做这个了,知识比较老化。
1.比较两个arm的sequence homology between 129 and B6.
一般的担忧是因为两个strain的序列有差别,可能影响同源重组的效率。但是如果幸运
的话也许你感兴趣的区域两者高度同源。
否则,几个OPTIONs
A。选用B6 derived ES CELL,有些service提供的。
B.让你的arm尽量长. 但是还是不能保证一定会有positive clone,要看运气,并且问题
是plasmid大到一定程度就不稳定了。比较safe的办法是用BAC recombineering做
BAC based construct.
C。 use long-range high fidelity DNA polymerase to PCR 129 genomic sequence.
这个对cloning要求有点高,并且要做sequencing确定没PCR error
n********k
发帖数: 2818
49
来自主题: Biology版 - clone求助,很特别的基因序列
1. synthesis or any cDNA clones outthere...or just synthesis the little
piece and then fusion PCR to the entire piece...
2. Primers ahead of the region
3. Primers after this region but with this long tails and go with two
step PCR: 94 68, also try Primestar,
4. PCR small fragments and then fusion PCR...
5. PCR other regions and then add this little piece through PCR-
mutagenesis...
6. Get a bac/genome clone for this gene and PCR in small pieces and
then Fusion PCR, or just PCR this hard region a
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