|
m****n
发帖数: 75 | 2 手上目前2篇稿需评审
1 是篇review
生物物理方面，单个分子的FRET在生物学中的应用
2 review
FRET在神经生物学中的应用
请站内信您的CV和联系方式 |
|
m****n
发帖数: 75 | 3 手上目前2篇稿需评审
1 是篇review
生物物理方面，单个分子的FRET在生物学中的应用
2 review （已经送出）
FRET在神经生物学中的应用
请站内信您的CV和联系方式 |
|
x*****o
发帖数: 441 | 4 你这个实验具体是想得出什么结论呢? 如果要求不高我们一般都不校正. 你是做single
molecule FRET 吗? 做这个的人从来不校正的说.
以前实验室做得bulk FRET的实验,就是简单的测Ratio A来校正,操作很简单.
Z.K. Majumdar, R.P. Hickerson, H.F. Noller and R.M. Clegg, Measurements of
Internal Distance Changes of the 30 S Ribosome Using FRET with Multiple
Donor–Acceptor Pairs: Quantitative Spetroscopic Methods, J. Mol. Biol (2005
) 351, 1123–1145. |
|
T****O
发帖数: 407 | 5 如果你经常测一些不太常见的荧光基团的激发－发射光谱，在操作上就不应该觉得FRET
很困难。
FRET效应可能非常微弱，对检测器的灵敏度和波长选择性要求都比较高。最好，先用一
个已知FRET效应的对子来校验你的仪器设备。 |
|
T****O
发帖数: 407 | 6 我不熟悉这样的仪器配置。
我熟悉的FRET对子是生物化学方面的。FRET产生条件相对苛刻，实际工作中，看不到
FRET往往倒是正常的。如果你的也是生物化学分子，有必要提醒一下，不要轻信你的结
果，不管是正结果还是负结果。多找几个条件，多找几个参照，而且要设计物理化学原
理不同的其他实验来印证。 |
|
f*********r
发帖数: 10 | 7 quench会改变fluorephore的quantum yield,如果要准确计算FRET你需要知道QY到底被
该变了多少。如果是定性估计，也许可以估计个QY的值。
Chem. Rev. 2006, 106, 1785-1813， 是个不错的review,公式12，13 calculate the
correction of FRET. |
|
z********u
发帖数: 94 | 8 做的是bulk FRET，因为我想比较两种情况下的区别，然后两个情况的quenching稍有区
别但是efficiency差别很大，所以想能不能correct quenching然后得到更确切的结论
。这篇paper已经读过了，他区别了dynamic quenching和static quenching，但是没有
具体给dynamic quenching的校正。 ratio A就是 FRET acceptor/direct acceptor对
吧，这个要怎么校正呢？
多谢多谢啊。
single
2005 |
|
a*******u
发帖数: 19 | 9 A-CFP和B-YFP（或B-CFP&A-YFP）共转HEK293T，COS7，使用lipofectamine 2000转染（
按说明书操作）48小时后，细胞中只有A-CFP或B-YFP，但是没有2个蛋白一起表达的细
胞。A在80 kD左右，B在90kD左右，是不是A和B蛋白太大，两个不容易一起表达？如果
将用A的剪切体约45kD再和B全长去做FRET估计能共表达吗？
Hela里共转Flag-A和His-myc-B的时候A表达很低，所以就没试；293T和COS7中Flag-A和
His-myc-B共转都没问题。 |
|
e***o
发帖数: 344 | 10 做蛋白互做检测时这2种方法，那种得灵敏度更高些？还是都不太靠铺？ Split GFP vs
FRET
谢谢。 |
|
I*****y
发帖数: 6402 | 11 都不太靠谱。
split gfp是说BiFC? 这个如果work了就work了，算你走运，如果不work，你连
troubleshooting都不知道咋下手。fret 相对还靠谱一点。
vs |
|
b******y
发帖数: 627 | 12 FRET is within 5nm. If your proteins are large, the chance of success is low
, too. |
|
e***o
发帖数: 344 | 13 I thought for FRET, it is 10 nm. How about split gfp/BiFC? Thanks！
People are using split GFP (called GRASP) to reconstruct pre/post synaptic
split GFP and get good signal. The distance of synapse is around 20-40nm.
The extracellular part of XFG isnt big. I am wondering 10nm is good for split gfp/BiFC?
low |
|
b******s
发帖数: 5365 | 14 不知道split GFP怎么做，FRET相当不好做，需要很仔细的control。 我们这里一个不
走运的博士生花了4年make it work。
vs |
|
C*******e
发帖数: 4348 | 15 考虑一个大的蛋白复合物
比如是正方体，边长大于50埃
考虑是实心的
那么顶面的donor有可能传递能量到底面的acceptor么？
donor和acceptor标记都在表面，不在蛋白复合物内部
还有一般而言FRET可以检测到的距离是在哪个范围内？
如果说数据是10%-30%的话 |
|
C*******e
发帖数: 4348 | 16 谢谢楼上两位的回复
不是我要设计实验
是在读一篇paper
然后我觉得他们的设计比较扯
但是缺乏相关知识
不清楚是自己太挑刺了还是他们的解释真的很牵强
我比较关心的是对于正方体顶面和底面之间
有可能FRET么？ |
|
z********u
发帖数: 94 | 17 1，如果你是指蛋白会不会影响fluorescence“传递”，我原来也问过系里的老师，他
的答案是蛋白相对于florescence是“transparent”的，所以底面顶面应该不会影响。
2，IAEDANS和5-IAF这一对的话，你可以查一下这对对应的r0，bulk FRET的话
efficiency大于0.1(OR even less)就可测量的了。不过附议楼上同学的意见，绝对值
没意义，还是要看变化。 |
|
m****n
发帖数: 75 | 18 一篇review，关于单个分子FRET
求审稿人
有兴趣的请站内信您的联系方式和学术背景
谢谢 |
|
BG
发帖数: 515 | 19 最近下载了Taekjip Ha group的软件处理FRET的数据。他的数据采集程序是C++平台的
，在Windows无法运行。他的网站提供的样品数据是.pma格式的。
不知道这里有人了解怎样生成这个.pma格式的文件？
我现在采集的数据是.tif格式的图片和录像。
非常感谢！ |
|
J*****i
发帖数: 77 | 20 请问做fret 需要用特殊的荧光仪吗，一般教学用的可以做吗？ |
|
J*****i
发帖数: 77 | 21 谢谢楼上，我试了一下，fret信号还挺明显的，只是看到有文献上提到用
phosphorimeter accessory,是不是用了它，信号会强很多，这个phosphorimeter是做
什么的？ |
|
A******y
发帖数: 2041 | 22 According to the specialist selling the instrument. To do Fret and
florescence polarization, it is better to have time-resolve capability. I
think you can do it, if the signal is good enough but that will effect your
range of sensitivity. |
|
m****n
发帖数: 75 | 23 一篇review，关于单个分子FRET
求审稿人
请站内信您的联系方式和学术背景 |
|
m****i
发帖数: 1076 | 24 Lesson: 3
Title: BLUE NOTES AND PASSING TONES, BENDING.
Style: Blues, theory
Level: Intermediate
Instructor: Olav Torvund
If you read my previous lesson on the blues scale, you will remember
that the blues-tonality is unstable and unsettled, because you play a
minor of scale over major chords. You should also remember that one and
a half steps up from the root gives you a minor third, and two steps up
from the root gives you a major third. I will also remind you of the
VII note, where you have t... 阅读全帖 |
|
n*********0
发帖数: 925 | 25 http://www.glorypress.com/devotional/FaithAndLifeOneYearBook.asp?bid=1
烦恼的罪
经文: 「不要….心怀不平。」（诗卅七1）
这是上帝的命令，和「不可偷盗」（出廿15）一样是上帝的命令。现在让我们
替「心怀不平」下一个定义。到底什么叫作「心怀不平」呢？在这里有一个很好的定义
：「『心怀不平』是平面上起皱纹。」「磨擦或者消磨」。一个乖戾的、无理性的、吹
毛求疵的人不但消磨自己，并且消磨别人。 「心怀不平」就是心中烦恼。这是有害的
，上帝不要我们自己伤害自己。
医生能告诉你：发一次怒，对于身体损害，比发一次热还厉害，所以一个常常
「心怀不平」的人不能得到康健的身体，因为我们知道「心怀不平」容易叫我们生气，
生气容易叫我们发怒。让我们服从上帝的命令──「不要….心怀不平！」──襥登（
Margaret Bottome）
Sin of Worry
Scripture: "Fret not" (Ps. 37:1).
This to me is a Divine command; ... 阅读全帖 |
|
m****i
发帖数: 1076 | 26 Lesson: 9
Title: Right hand Left hand technique - cont.
Level: Beginner
Style: Technique
Instructor: Tim Fullerton
This is part two in a series of how to develop good right and left
hand
technique for pick-style guitar.
*****THE LESSON****
PART II -- left hand position
This series is the approach that I use to teach pick-style
technique to all of my students. For best results, take these
articles to an educated and experienced teacher who is stylistically
broad based and who is acquainted with t... 阅读全帖 |
|
m****i
发帖数: 1076 | 27 Lesson: 2
Title: THE BLUES MELODY AND BLUES SCALE.
Style: Blues, theory
Level: Intermediate
Instructor: Olav Torvund
Those who want to play blues, should know the blues scale. Guitar
players should know how to finger blues scales all the way up the neck in the
so called "box position". But let us start with some peculiarities of
blues tonality.
If you start from the root note and play a scale, a major scale is
like this: (A major scale is often called a diatonic scale.)
I + II + III - IV + V + V... 阅读全帖 |
|
a*******l
发帖数: 2353 | 28
good!
still a little bit confusing . let me figure out.
if you acseding like position 1. from 1st string , 8 fret . then 1st string 5
fret ,then 2string 8 fret. how to keep your finger ? keep your index on 1st
string ,5 fret, then play 2st,8fret? How about on the same string?
that's a problem for me .
till now , my teacher told me to touch the board with pink around the end of
the second pickup.
I think he will probably tell me no need to touch it later. |
|
J**a
发帖数: 215 | 29 How about single molecule FRET? Steve Chu虽然不是发明FRET(Fluorescence
Resonance Energy Transfer)的人,但是第一个把它用于生物体系的单分子研究.你究竟看完了我的贴子么? 在FRET上,Steve刚好和optical tweezers的情形相反. 就我所知, FRET现在在
生物上的应用比optical tweezers还多,影响还大.包括现在基于single molecule
fluorescence技术上的super-resolution microscopy (STED, STORM, PALM). |
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p*l
发帖数: 1359 | 30 做个广告。
我做多色的FLIM-FRET，现在显微镜差不多好了，找了个合作准备做multi-color 3D
time lapse FRET看signal cascade。基本能用FRET成像的我都能做。果蝇，yeast，鱼
，还有细胞，能上显微镜台的都成。将来想做活老鼠的话，我也能自己做endoscope。 |
|
p******s
发帖数: 938 | 31 Sting给杀手里昂配的片尾曲，词曲精彩深刻不说，那原声吉它的演
奏实在是精彩之极。在网上找到几个版本大家看看：
Shape Of My Heart - transcribed by Luis Ferreira
This is NOT the way Dominic Miller plays it. I've got Sting's concert in Oslo
(1994) in videocassette and it is definitely harder than this. Dominic Miller
uses an electric-shaped acoustic guitar, which makes it easier to play in the
high frets as he does. The names of the chords might not be correct, too.
( If you want to play with the record, then use a capo on the 2nd fret )
Intro:
... 阅读全帖 |
|
m****i
发帖数: 1076 | 32 Lesson: 5
Title: The Pentatonic Scale
Level: Beginner - Intermediate
Style: Theory
Instructor: Bill Quinn
Hello, I'm Bill. I've thought a lot about the '1st lesson' but the
only thing that really makes sense is to just jump right in. For those
of
you who know the pentatonic scale - this will surely be a nice review,
for
the rest of you, get ready for the 1st step in playing lead.
The guitar makes learning scales easy! To most players, a scale is
just a memorized fingering pattern. Sure, the patt... 阅读全帖 |
|
a*******l
发帖数: 2353 | 33 来自主题: MusicPlayer版 - 单弦练习 这个单弦不是乐器，是吉它弦。
这个练习能够促进对吉它指板的理解，对音阶的连接把位的转换都很有帮助。 以前的帖
子似乎没提到过。
这个练习就是在一根弦上跑音阶。 我们可以从2弦(空弦b)C major开始.
C D E F G A B C D E
fret 1 3 5 6 8 10 12 13 15 17
练习举例，
上行
finger 1 2 4 1 3 4 1 2 4 1 2 4 1 2 4 1 3 4 etc
fret 1 3 5 3 5 6 5 6 8 6 8 10 8 10 12 10 12 13 etc
能跑多远跑多远，注意上行是用食指领头换把位。
下行
finger 4 3 1 4 2 1 4 2 1 4 2 1 4 3 1 4 2 1
fret 13 12 10 12 10 8 10 8 6 8 6 5 6 5 3 5 3 1
下行由小指领头。
上行比下行稍微容易些，因为平时练习时，食指的最低位置定义了把位，一想到把位就想
到 |
|
u***s
发帖数: 873 | 34 手感不错，容易弹。但 action 很低的时候，打品。
我去GC检查，他们说 Fret level有问题， 现在这把琴在prs的Maryland原
厂做Fret Level, 可能下个星期就可以送回来了。
在gc买的琴， 都可以让他们setup,
我新买的gibson sg special，在gc 做了Fret level， 然后setup 了几次，
终于达到了我的要求，action很低，又不打品(buzz). |
|
S*M
发帖数: 10832 | 35 谢谢，google了一圈，才知道什么意思
就是弦碰到fret吧
你这么一说，我才想起来我的琴可能真的打品
比如我用capo on 5th fret的话
后面的弦就非常非常低了，几乎碰到fret了
这个时候随便扫个什么和弦
声音往往特别muddy
有可能是打品了
郁闷
raise action是不是得花一笔钱啊 |
|
n*********0
发帖数: 925 | 36 http://www.glorypress.com/devotional/FaithAndLifeOneYearBook.asp?bid=1
不要忧虑
经文: 「应当一无挂虑。」（腓四6）
许多基督人不断地过着焦急挂虑的生活，有的竟烦恼得可怜。怎样能在日常忙
乱的生活中得到完全的安息是一个值得知道的秘诀。让我们先看：挂虑究竟有什么用处
？挂虑从来没有使谁强壮过；从来没有使谁遵行上帝旨意过；也从来没有使谁逃出困难
过。挂虑只会损害有用的生命、美丽的生命。焦急、挂虑、担忧，都是我们的主所绝对
禁止的，我们的主说：「不要….忧虑吃什么….穿什么。」（太六25）主的意思并不是
说：我们用不着预先思想，我们的生活可以毫无计划、毫无规律；祂乃是说：我们用不
着为这些事挂虑。你以为你这样挂虑、担忧，别人不会知道么？我告诉你：从你脸上的
愁纹，从你说话的声调，从你生活中最微小的动作，从你缺乏喜乐的精神看来，别人早
已知道了。我们应当把我们的挂虑卸给上帝！因为祂说：「凡劳苦担重担的人可以到我
这里来，我就使你们得安息。」（太十一28）──沙勤德（Rev. Darlow Sargeant）... 阅读全帖 |
|
p*l
发帖数: 1359 | 37 there is no way to do a four part complex FRET as of now. It is an optical
instrument research problem. Current
microscopes just can't give you enough information to analyze FRET between
so many labels. You can try one pair
FRET at a time. But again what you get are not direct evidences of a four
part complex. |
|
b******s
发帖数: 5365 | 38 很精彩的讨论啊，学习了！谢谢各位牛牛！
Sunnyday: 知道了cytosolic protein B和membrane protein A的binding region的情
况下，那么用tagged B binding region应该可以克服B和其他蛋白作用而导致IP A信号
弱的问题了吧？这个和你说的tagged A fragment IP B是一样的，可以算个反向IP.
关于FRET，这个能做出来当然非常好，而且可以证明是直接binding了，不过我对这个
assay一直是心存敬畏啊。如果已经有optimised 好的系统那还好，要是要从头
optimise的话，可能会很悲催。我知道有个phd student 花了4年时间来做FRET,文章是
发了，通篇就是FRET，但觉得很risky啊。 |
|
w*******y
发帖数: 60932 | 39 World Music Supply has the Squier Classic Vibe 50's Strat Lake Placid Blue
for $242.99 with free shipping. No CA tax! Must add to cart to see final
discount.
Next cheapest I could find is 379 from Sweewater. I also think this is a
better deal than a MIM Strat of Craigslist. At least in SoCal, the post-2006
Strats go over $250 - plus, you know... they're used.
Link:
http://www.worldmusicsupply.com/squier-classic-vibe-50s-strat-l
{IMGhttp://www.worldmusicsupply .com/zoom_img/_1241080549.jpg
... 阅读全帖 |
|
s******r
发帖数: 5309 | 40 关税导致通胀预期增加，迫使美联储加息是这次股灾的原因。
President Donald Trump during a news conference in the Rose Garden of the
White House in Washington, October 1, 2018.
Kevin Lamarque | Reuters
President Donald Trump during a news conference in the Rose Garden of the
White House in Washington, October 1, 2018.
While President Donald Trump has directed his ire at the Federal Reserve for
the recent stock market decline, he also has himself to blame.
The president's escalating trade battle with China has added another la... 阅读全帖 |
|
c*****g
发帖数: 21627 | 41 Excellent article
英文原文：
Barron's Cover | MONDAY, JANUARY 28, 2013
The Next Boom
By KOPIN TAN
Cheap natural gas and increasingly competitive labor costs are bringing
factories and jobs back to the U.S. Eight ways to win.
As the only industrialized superpower not decimated by World War II, the
United States once made nearly 40% of the planet's goods. These days, that
number has shrunk to 18%. We make American Girl dolls in China, Levi's jeans
in Mexico, and enough movies in Vancouver to nickname ... 阅读全帖 |
|
l****z
发帖数: 29846 | 42 CNN's Blitzer Frets Israelis Carrying Guns to Defend from Palestinian
Stabbing Attacks
By Brad Wilmouth | October 14, 2015 | 11:32 PM EDT
As he interviewed Israeli police spokesman Micky Rosenfeld on Wednesday's
The Situation Room, CNN host Wolf Blitzer repeatedly showed concerns that
Israelis may be doing too much to defend themselves amid a spate of
stabbings against Israeli Jews by Palestinians in Jerusalem.
All five questions posed to the police spokesman had the premise that
Israelis are go... 阅读全帖 |
|
g********2
发帖数: 6571 | 43 It’s time for Hillary Clinton to start worrying about her Florida campaign.
Despite roughly 500 full time campaign staffers spread across Florida to
help Clinton win forever close Florida, party leaders and activists across
the state are fretting about the campaign’s ground game. Even Clinton
campaign staffers in other states are quietly buzzing about the Florida
campaign struggling to meet its goals for voter registration and other
outreach benchmarks.
...
Some of the raw numbers in Florida are... 阅读全帖 |
|
|
n******8
发帖数: 558 | 45 Harp
I erect a harp by the mirror lake.
In the mirror I gaze at her grace,
and wish she would sing when awake.
When autumn breeze wrinkles her face,
I hear fallen leaves pluck her string.
And see golden sun rays decorate her reflection.
Harp sings a song of mesmerizing spring,
Till I tie autumn with affection.
竖琴是很有意思的乐器。 几天前开车带小宝去上游泳课， 电台播放着很好听的音乐。
我是家里公认的乐盲， 五音不全， 对音乐的感知仅限于好听和不好听的欣赏水准。
因为好听， 就问小宝， 是什么乐器。小宝答， 竖琴。 我也不知哪来的信心， 坚定
地否决他， 是吉他， 想是因为大学被迫听过太多的吉他， 初学者的吉他。 痛苦的记
忆总是留的久些。 小宝自然不买我的。 好在音乐不长，... 阅读全帖 |
|
p******s
发帖数: 938 | 46 Sting给杀手里昂配的片尾曲，词曲精彩深刻不说，那原声吉它的演
奏实在是精彩之极。在网上找到几个版本大家看看：
Shape Of My Heart - transcribed by Luis Ferreira
This is NOT the way Dominic Miller plays it. I've got Sting's concert in Oslo
(1994) in videocassette and it is definitely harder than this. Dominic Miller
uses an electric-shaped acoustic guitar, which makes it easier to play in the
high frets as he does. The names of the chords might not be correct, too.
( If you want to play with the record, then use a capo on the 2nd fret )
Intro:
|
|
m****i
发帖数: 1076 | 47 Lesson: 8
Title: Development of Chords from Scale Tones in Thirds
Level: Beginner
Style: Chord theory
Instructor: Dennis O'Neill
10 February 1993
Introduction.
Many chords can be developed by extracting alternate scale tones,
i.e., using tones that are major thirds or minor thirds apart within a
scale. In this series of exercises, you will begin by building
three-note chords from the major and three minor scales relative to C
major and progress to building seven-note (thirteenth) chords. You
wil... 阅读全帖 |
|
g*******a
发帖数: 1383 | 48 i'd spend more money on a good amp, i realized that after several
years playing guitar. i have a line 6 and floor padel, but most
of time i don't use them. i got this cheapest marshall and an
ibanez lead padel, that's all i need.
RG is a great gutiar, jumbo fret means the distance between frets
are large. i think you'd better start w/ an epiphone les paul.
strat is not designed for distortion sound. les paul can do more
in that sense. esp. when you don't have a style yet.
wouldn't be a bad idea |
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t****d
发帖数: 127 | 49 I think everybody should learn how to read sheet music.. and some basic
theories.. I think fretboard logic is sort of a tricky topic.. check out that
mick goodrick book that I mentioned.. basically, there is no set way of how to
divide up the fretboard.. however u should know what each note is.. and
relative intervals.. for example, the interval between a note on a string and
the note on the same fret on the next higher up string.. is a perfect 4th.. ur
major third is one fret down on that highe |
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a*t
发帖数: 2596 | 50 ☆─────────────────────────────────────☆
ablution (菜鸟...学习中) 于 (Sun Jul 1 16:47:36 2007) 提到:
三个手指挤在一格
五弦老是有破音
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antiwheel (重振旗鼓， 大劈！) 于 (Mon Jul 2 08:49:52 2007) 提到:
355533?
other than using all 4 fingers, it also could be bared with your Ring finger.
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ablution (菜鸟...学习中) 于 (Mon Jul 2 09:47:22 2007) 提到:
yes 355533 or Asus 577755 etc
3rd fret use index finger?
5th fret use ring finger?
then 1st and 2 |
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