w*******y
发帖数: 60932 | 1 Through Sunday night at 9:59 EST (or while it lasts), you can order one year
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w*******y
发帖数: 60932 | 3 Click here for the deal!:
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coupon is: GOLF
Golf Digest is edited for golf enthusiasts at all levels of ability-from
beginners to low-handicap players-to help them improve their play and enjoy
the game even more. Each month, Golf Digest provides in-depth, "how-to"
instruction and tips by an unparalleled team of teaching professionals and
playing editors, including major championship winners like Ti... 阅读全帖 |
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X***h
发帖数: 74 | 4 如果我PCR后的product比较干净，能否不处理直接restriction digest? 现在我比较麻
烦的是如
果用column purification总是丢失大部分的DNA，后面digest完了还要gel
purification，
那就所剩无几了。
或者用SAP 去除primers，然后Ethanol-sodium acetate precipitation，总可以上
digestion了吧。 |
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s******r
发帖数: 2876 | 5 你既然都P出来了，完全可以用PCR产物第二次扩增一遍，
DNA还不是要多少有多少。
如果我PCR后的product比较干净，能否不处理直接restriction digest? 现在我比较麻
烦的是如
果用column purification总是丢失大部分的DNA，后面digest完了还要gel
purification，
那就所剩无几了。
或者用SAP 去除primers，然后Ethanol-sodium acetate precipitation，总可以上
digestion了吧。 |
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S******e
发帖数: 393 | 6 想用SalI-NheI double digetion,
NEB suggest sequential,
但我看到SalI和EcoRI double digestion 用 E.coRI buffer
NheI 和 EcoRI double digestion 也用E.coRI buffer
是不是SalI-NheI double digestion 就可以用E.coRI buffer，
而不用sequential了？
多谢回答！ |
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e******0
发帖数: 2149 | 7 现在在用LC/MS/MS做一个膜蛋白的定量，试过run western blots的同时做in gel
digestion 然后LCMSMS，也试过in solution digestion.听别人说过似乎in gel
digestion比in solution的效率高，但我感觉in gel technically 难做些。至今两个
方法做出来的结果差别不大。想听下前辈们的意见，哪个方法会好些。多谢！ |
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w*******y
发帖数: 60932 | 8 LINK:
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refresher on fundamentals, and Pocket Tips, a review of the issue's best
tips. You'll also get equipment reviews, Ame... 阅读全帖 |
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c***e
发帖数: 111 | 19 不知道该发哪里，就来这里求助大家了。n个月前，在Reader's Digest上Enter了一个
抽奖，结果没注意他下面有个小小的checkbox订阅他们的杂志，我没有主动uncheck，
于是就变成说我订阅了。那个抽奖只要enter就送一本免费的cooking书，我当时住学校
，因为我不想订阅杂志，那本cooking的书我根本就没收，我要求学校的宿管收件处
return了。后来也从来没有收到过任何一期杂志，第一次寄来bill我没理，12月第二次
催我要钱，说晚交有late fee，我又没理。我觉得我没有收到杂志，也没想订杂志，不
用理他。现在又发来最后一封催钱的信。警告我再不交订阅费+late fee，就要找第三
方independent collection agency来讨债。
我在杂志上留了学校的名称和自己的真名，它们一直在往我学校的宿舍地址寄，我现在
已经搬走了，我该怎么办呢。40刀钱不至于把我告上法庭吧。
查了网上说Reader‘s Digest经常莫名其妙寄东西来要钱，我怕我的支票开出去，以后
它自说自话来划钱了。而且我根本没想订杂志（虽然事实是我没有uncheck那个b... 阅读全帖 |
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D*a
发帖数: 6830 | 20 Quick Tail Prep
Collect 1 mg skin biopsy (= normal size tail or figure tip) in 0.5 ml
Eppendorf tube.
Add 100 microliter of quick tail digestion buffer (below).
Incubate @ 56 C several hours to overnight.
Vortex and incubate @ 99 C, 10 minutes.
Centrifuge @ max speed, 5 minutes.
(NB i don't vortex so i don't need to centifuge the tube, it works)
Use 1-2 microliter aliquot of supernatant for a 25-50 microliter PCR
reaction (or whatever you routinely do).
Transfer supernatant to a fresh tube for s... 阅读全帖 |
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D*a
发帖数: 6830 | 21 Quick Tail Prep
Collect 1 mg skin biopsy (= normal size tail or figure tip) in 0.5 ml
Eppendorf tube.
Add 100 microliter of quick tail digestion buffer (below).
Incubate @ 56 C several hours to overnight.
Vortex and incubate @ 99 C, 10 minutes.
Centrifuge @ max speed, 5 minutes.
(NB i don't vortex so i don't need to centifuge the tube, it works)
Use 1-2 microliter aliquot of supernatant for a 25-50 microliter PCR
reaction (or whatever you routinely do).
Transfer supernatant to a fresh tube for s... 阅读全帖 |
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y****6
发帖数: 196 | 22 Thanks for sinister and kaka1000's reply. I labeled the antibody with
fluorescent dyes and quenchers, and I want to digest the antibody to small
fragments to break the FRET between dyes and quenchers. It would be best if
I can digest the antibody into the smallest fragments. Papain may not be the
best choice as it only cut antibody into three pieces. Maybe I can use some
kind of protease mixture? Any suggestion is welcome. |
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s******s
发帖数: 1584 | 23 两种Digestion没区别 要是担心Digestion 不完全多加点Trypsin就好了
膜蛋白我推荐In Gel |
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f******h
发帖数: 609 | 24 5个包子答谢:求一篇 2009 IEEE intermag conference 的 digest
找不到会议的CD了, 哪位仁兄还有的话能否帮我email一份? Please note it's not the paper. I have the 6page paper. I want the 2page digest from the conference CD.
FD-02 Spin polarized transport and dynamics in magnetic tunneling structures.
万分感激! 我的地址: h******[email protected] |
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f******h
发帖数: 609 | 25 5个包子答谢:求一篇 2009 IEEE intermag conference 的 digest
找不到会议的CD了, 哪位仁兄还有的话能否帮我email一份? Please note it's not the paper. I have the 6page paper. I want the 2page digest from the conference CD.
FD-02 Spin polarized transport and dynamics in magnetic tunneling structures.
万分感激! 我的地址: h******[email protected] |
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w*******y
发帖数: 60932 | 31 Link:
http://www.discountmags.com/
Maximum PC for $7.99/year
Maxim for $3.99/year
Wired for $3.99/year
Reader's Digest for $3.99/year
Code
READ2011
This is a great deal on Maximum PC and Reader's Digest
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w*******y
发帖数: 60932 | 34 Tanga
Large Print Readers Digest Magazine Subscription
$24.99 - $21.49 code DIGEST
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w*******y
发帖数: 60932 | 35 Tanga
Readers Digest Magazine Subscription
$24.99 - $21 code DIGEST
= $3.99
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w*******y
发帖数: 60932 | 36 Tanga
Readers Digest Magazine Subscription
$24.99 - $21.49 code DIGEST
= $3.50
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w*******y
发帖数: 60932 | 39 Readers Digest $2 per year with coupon code RD128 from publicationstation.
http://www.publicationstati on.net/Readers-Digest-11-issuesyr_p_98.html
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x****5
发帖数: 1564 | 41 1. get your stance in "sink"
2.put your takeaway on slow-mo
3. stop your backswing before you start down
4.make a "one-piece" downswing
5. anchor your right foot
sorry it is in the current golf magazine, not golf digest |
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d****9
发帖数: 3206 | 42 http://www.discountmags.com/
Readers Digest Magazine Subscription for $2.99 (up to 4 years)
coupon code 3051
我刚定了两年的，这么便宜，学学英语吧。。。。 |
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x******r
发帖数: 367 | 43 Hello!
Is there anywhere that I can buy Readers Digest and Times in China?
Thank you very much. |
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a********a
发帖数: 3176 | 44 Reader's Digest has been out of business. |
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x******r
发帖数: 367 | 45 I am just asking if it is possible to get Readers Digest issues after 2000,
not necessarily the most current one.
Thanks. |
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l***r
发帖数: 459 | 46 I met a strange problem on JDBCRealm:
In my server.xml, it looks like:
className="org.apache.catalina.realm.JDBCRealm"
debug="99"
driverName="com.mysql.jdbc.Driver"
connectionURL="jdbc:mysql://localhost/auth"
connectionName="user"
connectionPassword="password"
userTable="users"
userNameCol="user_name"
userCredCol="user_pass"
userRoleTable="user_roles"
roleNameCol="role_name"
digest="MD5"
/>
It works if I use FORM authentication. So I am sure
mysql driver and MD5 are working fine. But it d |
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M******t
发帖数: 555 | 47 digest 100ug plasmid DNA 最小体积多少啊?
谢谢. |
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m******5
发帖数: 1383 | 48 google N-chip和X-chip
两种不同的方法
用酶digest的非特异性binding少，并且因为去掉了cross linking的步骤，能用的抗体
数大大增加。缺点是很难拿到transcriptional factor和特定片断的binding 。做
Histon的人应该用得多些，但因为没有cross linking，也会引入很多非特异性的
nucleosome remodeling。
用sonication 以及cross linking配合的方法好处是拿到的相互作用理论上说更接近生
理条件，但因为引入了固定这个过程，也导致许多artificial 作用， 比如蛋白质，
DNA间非功能性的相互作用 |
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s******u
发帖数: 81 | 49 多谢！
另外的原因可能还有digestion enzyme的sequence preference.
说到co-IP, lysis buffer一般都加detergent吗？看别人的文章基本都加，可有的
interaction加了detergent几乎检测不到。
stringency |
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m******t
发帖数: 109 | 50 i want to know the digestion buffer for genotyping that doesnot purify
genomic DNA.i forgot it. let me know. thx. |
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