e******e
发帖数: 28 | 1 大家有人有在sonic automotive dealer买车的经验吗？他们的sonic price很低，而且
一口价，没有negotiation. 我家因为两地 (LD 在TN， 我在NC)，LD工作的那边有
sonic dealer，给的honda odyssey EX-L 才30792 (不包括TTL)，而我这边是普通
dealer，他们给的internet quote都不低于33000TTL（不含TTL）, 我还没有去dealer
那里砍价。所以我们想有没有可能在LD那（Tennessee）的sonic dealer那里买车，并
交我在的州（NC）的tax,上NC的tag. 然后把车开回NC。请问这样可行吗？
sonic dealer的车有什么不同吗？ 有什么注意的地方吗？ |
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e******e
发帖数: 28 | 2 大家有人有在sonic automotive dealer买车的经验吗？他们的sonic price很低，而且
一口价，没有negotiation. 我家因为两地 (LD 在TN， 我在NC)，LD工作的那边有
sonic dealer，给的honda odyssey EX-L 才30792 (不包括TTL)，而我这边是普通
dealer，他们给的internet quote都不低于33000TTL（不含TTL）, 我还没有去dealer
那里砍价。所以我们想有没有可能在LD那（Tennessee）的sonic dealer那里买车，并
交我在的州（NC）的tax,上NC的tag. 然后把车开回NC。请问这样可行吗？
sonic dealer的车有什么不同吗？ 有什么注意的地方吗？ |
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j****t
发帖数: 1663 | 3 我的sonication 步骤如下，具体输出功率不详（可以参考Branson的说明书）。0.8%
SDS 应该足够强了.我用的是0.1% of Na-deoxycholate and 0.2% of sarkosyl （以前
用过0.5% of sarkosyl）。我有时候看lysate是否clear了，来决定sonication的次数。
Sonicate sample (in a 15 ml falcon tube) using Branson 450 sonicator (Power
Set 2 or 4, duty cycle 100%, 4-6 times, 30 sec each with 1 min interval).
During the sonication, keep falcon tube in a 50 ml beaker filled with ice. |
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w*******y
发帖数: 60932 | 4 Street Fighter 4::
http://itunes.apple.com/us/app/street-fighter-iv/id354655665?mt
.99
Sonic the Hedgehog: 1.99
Sonic the Hedgehog 2: 2.99
Sonic the Hedgehog 4 Episode 1: 3.99
Sonic Spinball: .99
100% of Sega's proceeds from Sonic the Hedgehog iOS titles will be donated
to the red cross disaster relief efforts in Japan.
Capcom will also donate all the sales of Street Fighter 4 to public charity
from March 15th through March 22nd.
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w*******y
发帖数: 60932 | 6 Did a search and didn't find anything about it.
Bay Area local ISP Sonic.net has dropped their "Fusion" service (high speed
DSL + land line) from 50/mo to 40/mo. Speeds are 20down/1up. Unlimited
nationwide calling, free 3 way calling, free caller ID, free call waiting (
features that AT&T made me pay for).
Sonic.net also has award winning customer support and is routinely ranked as
one of the Bay's best ISP.
Not trying to make it sound like an ad but Sonic has been absolutely awesome
. I was so ... 阅读全帖 |
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b*******1
发帖数: 1352 | 8 正在att, comcast和sonic间犹豫。
sonic看起来不错，但家里据中心7000 feet，网速会下降很多吗？另外请问sonic在下
订单后多久能把网装好，网络稳定吗？
先谢过了。 |
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x*******9
发帖数: 12 | 9 朋友送了套Clarisonic Mia On-The-Go Sonic Skin Cleansing System， 里面有
Clarisonic Mia cleansing unit and pLink charger
One sensitive skin brush
One normal skin brush
1 fl oz gentle hydro cleanser
1 fl oz nourishing care cleanser
1 fl oz refreshing gel cleanser
Brush heads (2) precision-designed to oscillate bi-directionally at sonic
frequency
因为我已经有一套了，所以这套（new in box）想转让， 有意者站内联系吧
Clarisonic Mia On-The-Go Sonic Skin Cleansing System |
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w********u
发帖数: 13 | 10 谁来说说Sonic的牙刷到底比普通电动牙刷好在哪了？然后Sonic和Sonic之间又有什么
区别？那些Fancy的功能到底有没有用？我看现在飞利浦家最好的是HX9332，还能美白+
抛光，感觉都不用去牙医那洗牙了~ |
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j*****a
发帖数: 658 | 11 Don't quite understand your question.... You want to detect the
phosphorylation of a protein, why not use western? Even if you do ELISA to
see p-protein, no need to sonicate....RIPA is strong enough to get all your
cytosolic and nuclear protein out. Sonication mostly is used to break down
chromatin, DNA or chop down plasma membrane to release cytoplasma membrane
protein....If I am wrong, please someone corrects me.. So I think in your
case, you probably don't need sonication at all.
Also, freez... 阅读全帖 |
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j****t
发帖数: 1663 | 12 我是在这个lysis buffer里作sonication的。我现在用0.2% sarkosyl（instead of 0.
5%）,和 1% of Na-deoxycholate。sonication的效果很好。而且这个浓度的detegent
对于IP 来说还是高了些，我会在做IP前把sample稀释一倍。
我觉得你的lysis和sonication 的条件也许可以了，问题说不定是在reverse-
crosslink。我的快速检测片段大小和ChIP DNA浓度的步骤如下，供你参考参考。
Check chromatin size and conc.
1. Dilute 50 ul chromatin extracts with 50 ul TE buffer. Add 4 ul of 5 M
NaCl and 1 ul of 10 mg/ml Proteinase K (Invitrogen #25530-015) (use a PCR
tube).
2. Incubate at 65 oC for at lease 4 hr (using a PCR machine) ... 阅读全帖 |
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H****n
发帖数: 26 | 13 最近做一个Transcription Factor（120KD）的ChIP， 很奇怪的事情，--
前面标准的1% formaldyhe RT 10min，0.125M Glycine RT 5 min, PBS wash,
裂解完细胞后，温和的sonication完后，input用western看发现原来条带的位置没有（
或者变的很淡），在下面有两条小的蛋白带。 做了一下RNA pol-II的WB (>150kD <
200kD)也是类似的情况，原来条带的位置淡了很多，在底下出现了两三条size小的条带。
先后试了RIPA (0.1%SDS)和nuclear lysate(1%SDS)裂解，效果一样。只要有
sonication,即使是很温和的5个cycle 30s/30s (该条件下DNA shear都不完全200-
3000bp), 原来蛋白条带就会消失，取而代之的是底下出现的size小的条带。
请问这是sonication 导致大分子量的蛋白容易降解吗？要如何解决？ |
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C******g
发帖数: 2930 | 14 Tired of making replica 1-1 levels from Super Mario Bros. in LittleBigPlanet
? Well, it's time to explore the other retro gaming classics. Sonic the
Hedgehog is coming to LittleBigPlanet this week with a new sticker and
costume pack for $5.99. Unfortunately, this isn't a level pack akin to Metal
Gear Solid 4 or Pirates of the Caribbean, so don't expect any high-speed
game modifiers. Instead, this pack is solely for Sonic fans that want to
play as Sonic, Tails, Knuckles, Dr. Robotnik, and yes ... |
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w*******y
发帖数: 60932 | 15 BestBuy
Sonic & Sega All-Stars Racing With Banjo-Kazooie (Xbox 360)
Sonic & Sega All-Stars Racing (PS3)
Sonic & Sega All-Stars Racing (Wii)
$19.99 with free store pickup, otherwise shipping is $2.49
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w*******y
发帖数: 60932 | 17 To go along with the Sonicare brush deals @ Amazon, I found this cover to
prevent moisture in the cavity.
Sonic Seal:
http://thesonicseal.com
Promotion:
like our fan page and well give you $5 off! Once you show up on our list,
your order will ship within 2 days. This is our new way of advertisingwe pay
our customers and not a marketing firm. Use the discount code sonicseal50
upon checkout. Enjoy your Sonic Seals!
Normal price is $10.99.
Coupon code is really 50% off and it worked for qty of 2.
S... 阅读全帖 |
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w*******y
发帖数: 60932 | 18 Since Dane is unable to promote his own company here by starting a new
thread on it, I figured I'd do him an unasked-for favor and promote his
company anyway, as a happy customer for several months now.
NEW RATES FOR SONIC.NET FUSION SERVICE:
$39.95/month for single-line ADSL (same price as Dec. 2010; no price drop)
$69.95/month for two-line Fusion residential (down from $79.95)
$49.95/month or $89.95 month for small business (single line vs. two-line;
old price was $60 and $120, respectively)
W... 阅读全帖 |
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w*******y
发帖数: 60932 | 20 So Sonic has daily specials all winter long. All 99 cents.
Monday- Chicken Strip Sandwich
Tuesday- 1/4 lb Sonic Burger
Wednesday- Corn Dog and Small Tot
Thursday- 6" Premium Beef Hot Dog
Friday- Grilled Cheese or BLT
Saturday- Mozzarella Sticks (5)
Sunday- Jr Sonic Burger and Small Tot
Thought this was a pretty good deal.
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l****u
发帖数: 1764 | 22 在costco 租车，选了个Enterprise家的intermediate， 上面写的Hyndai Elantra or
similar，到了机场我能选的只有一辆chevy sonic，然后一个劲叫我加10.99 upgrade
。后来没升，就开的sonic，感觉特别小，回来一查发现是subcompact，是不是被坑了
啊？应该是intermediate > compact > subcompact吧？一般都是免费upgrade，这莫名
其妙被downgrade两级，能不能投诉丫啊？
是不是costco的租车比较underestimate？（我之前在hotwire上租车compact的车都给
过我hyndai sonata，一般属于full size sedan了） |
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J**S
发帖数: 25790 | 23 现在用ATT DSL PRO +Phone 最低服务，网速速度慢，还要最后$48.换SONIC，值不值啊？
抢OO，抢AMEX WISHLIST无一收获，是不是该换了？
Sonic.net 20Mbps internet + unlimited phone service $39.95/mo |
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x*******9
发帖数: 12 | 25 朋友送了套Clarisonic Mia On-The-Go Sonic Skin Cleansing System， 里面有
Clarisonic Mia cleansing unit and pLink charger
One sensitive skin brush
One normal skin brush
1 fl oz gentle hydro cleanser
1 fl oz nourishing care cleanser
1 fl oz refreshing gel cleanser
Brush heads (2) precision-designed to oscillate bi-directionally at sonic
frequency
因为我已经有一套了，所以这套（new in box）想转让， 有意者站内联系吧 |
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x*******9
发帖数: 12 | 26 朋友送了套Clarisonic Mia On-The-Go Sonic Skin Cleansing System， 里面有
Clarisonic Mia cleansing unit and pLink charger
One sensitive skin brush
One normal skin brush
1 fl oz gentle hydro cleanser
1 fl oz nourishing care cleanser
1 fl oz refreshing gel cleanser
Brush heads (2) precision-designed to oscillate bi-directionally at sonic
frequency
因为我已经有一套了，所以这套（new in box）想转让， 有意者站内联系吧 |
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x*******9
发帖数: 12 | 27 朋友送了套Clarisonic Mia On-The-Go Sonic Skin Cleansing System， 里面有
Clarisonic Mia cleansing unit and pLink charger
One sensitive skin brush
One normal skin brush
1 fl oz gentle hydro cleanser
1 fl oz nourishing care cleanser
1 fl oz refreshing gel cleanser
Brush heads (2) precision-designed to oscillate bi-directionally at sonic
frequency
因为我已经有一套了，所以这套（new in box）想转让， 有意者站内联系吧 |
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J**S
发帖数: 25790 | 28 现在用ATT DSL PRO +Phone 最低服务，网速速度慢，还要最后$48.换SONIC，值不值啊？
Sonic.net 20Mbps internet + unlimited phone service $39.95/mo |
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h***i
发帖数: 427 | 29 看PPLIVE, PPSTREAM可否？
$19.95/month
Features:
Download Speed: 3.0mbps - 6.0mbps
Upload Speed: 512kbps - 768kbps
IP Addresses: 1 Dynamic (More Info)
Email Accounts: 15
Per Email Account storage: 1GB
Free Mailing Lists (More Info)
Configurable Firewall (More Info)
Advanced Anti-spam tools (More Info)
Multicast Access (More Info)
VPN Termination (More Info)
Disk Storage: 250MB
Hosted Bandwidth: 1GB
http://login.users.sonic.net/ (More Info)
Shell Access: shell.sonic.net (More Info)
Dialup Internet Acc... 阅读全帖 |
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s*******g
发帖数: 1607 | 30 why do people open their big mouth without knowing what the
hell they are talking about??? huh??
sonics has been struggling this year because baker is bothered
by the trade rumors in the offseason, and patrick ewing is
too old and lost the fire he once had. baker is upset at
westphal, so's payton for trying to trade his best buddy
baker is still an awesome player, he could've easily been
an allstar. westphaldoesn't have the personality to control
the sonics, he belongs to the old school of nic |
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m***8
发帖数: 7797 | 31 看到sonic,还以为是sonic drive in 呢。 |
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r*********e
发帖数: 29495 | 32 [参加活动]那支释去的超音速Sonics球队
曾经到主场看过3次超音速的比赛，2008年3月这次，算是它最终离开西雅图，搬到OKC
之前不久的一场比赛，所以也用此文，纪念那支已经不存在的超音速篮球队。
主要这次是当地校友搞活动，20块钱一张票，第16排，算是很好的deal，还包括7块钱
的餐卷。定了两张，和LD一起去看，比赛双方是超音速和开拓者，因为两个城市距离很
近，这算是西北地区主要的rivalry game了。到目前为止已经在这里看过两次NBA了，
这次也算是三进Key Arena了，之前两次都是有姚明领衔的火箭来访时的比赛，基本上
每次来，搞得都好像火箭主场似的。
KeyArena。西雅图的主场在NBA的声誉不是太好，硬件设施老化，主场座位不足，而且
，没有相应数量的停车场，直接对球市造成影响。
下午早早下了班，来到体育场，首先吃了垃圾晚饭，用餐卷买了热狗加可乐。之后来到
我们的座位，真不错，距离真近，连队员脸上的表情都看得很清楚，各种中场表演也是
看得更加真切。我们的周围有不少校友，大家聊了聊天，也是挺高兴。
关于比赛，今天超音速打得不错，最后时刻反超，获得了久违的胜利。当时... 阅读全帖 |
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x*******9
发帖数: 12 | 33 朋友送了一套Clarisonic Mia On-The-Go Sonic Skin Cleansing System， 里面有
Clarisonic Mia cleansing unit and pLink charger
One sensitive skin brush
One normal skin brush
1 fl oz gentle hydro cleanser
1 fl oz nourishing care cleanser
1 fl oz refreshing gel cleanser
Brush heads (2) precision-designed to oscillate bi-directionally at sonic
frequency
因为我已经有一套了，所以这套（new in box）想转让。有意者站内联系吧 |
|
x*******9
发帖数: 12 | 34 朋友送了套Clarisonic Mia On-The-Go Sonic Skin Cleansing System， 里面有
Clarisonic Mia cleansing unit and pLink charger
One sensitive skin brush
One normal skin brush
1 fl oz gentle hydro cleanser
1 fl oz nourishing care cleanser
1 fl oz refreshing gel cleanser
Brush heads (2) precision-designed to oscillate bi-directionally at sonic
frequency
因为我已经有一套了，所以这套（new in box）想转让， ask for $120.有意者站内联
系吧 |
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i******m
发帖数: 495 | 36 要做一个ELISA检测细胞膜上一个蛋白的phosphorylation. 按照protocol应该把细胞
lysis以后做sonication, 但是实验室没有sonicator, 有什么什么其他替代方法呢?
Freeze-thaw会不会把磷酸化给弄没了?
thank you |
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s******s
发帖数: 13035 | 37 MNase也可以先crosslink，甚至可以有人crosslink->MNase->sonicate
repressive marker貌似sonicate很难做好，用MNase能做的很漂亮 |
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n******2
发帖数: 971 | 38 No, my sonicating buffer is slightly different from lysis buffer.
They both contain low concentration of Tris, NaCl, MgCl2, CaCl2, however,
sonicating buffer consists of higher amount of IGEPAL4% (compare to 0.5% in
lysis B) and 0.8% SDS.
I'm following the protocol developed by Affymatrix. It worked well on mouse
cells performed by another lab, but bothers me very much on my human cell
line. |
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k*****n
发帖数: 323 | 39 还有一个，可以考虑稀释一下样品在做sonication，问一下你们sonicate用的什么机器
？ |
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