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T*******n
发帖数: 493 | 2 Try \caption[{Conc [a]}]{The concentration is [a]}
LaTeX (actually TeX) only pairs { with }. It doesn't know how to
pair [ with ] because it regards [ ] as punctuations, not grouping
delimiters. If you want to understand the details, you need to
know what catcodes are in TeX and how macros argument lists
are parsed. |
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r**r
发帖数: 171 | 3 A good CSF extract should be arrested @ metaphase.
After the addition of calcium or fertilization which causes a transient
increase in cytoplasmic calcium conc., the CSF will be inactivated, following
by APC activation, cyclin B destruction and mitotic exit. And this process is
roughly 30 min. It is easy to collect phosphorylated separase before the
addition of calcium. To collect the unphosphorylated one, like you say, you
can throw sperm chromatin into extract and monitor the progresstion by D |
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g****r
发帖数: 14 | 4 I have been using invitrogen 2 step RT kit for a while,
always give me very good result ...
On possible is in your case, your total RNA might
be partially degraded, so that you can only get the
shorter segment, instead of the 1kb product
suggestions:
1) double check your primer design
and PCR condition (TM, Mg conc. pfx? or taq, or pfu?)
2) if everything seems to be correct
be more strict with your RNA extraction
use the ambion RNaseZap to remove environmental factors
(just as much a |
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R****e
发帖数: 53 | 5 how do you tell which is or which is not bg for Tuj1?
It is a very good ab. Try low conc. |
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s******e
发帖数: 370 | 6 this shouldn't be very hard, tuj1 is one of the best antibodies
we order it from covance
also check other conditions as well, like 1st/2nd ab conc, species
reactivity, and so on. and have a positive control. |
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w******e
发帖数: 1187 | 7 多谢啦！我也确实偷懒，担心low conc.的gel太软就没折腾。。。
8% |
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w******e
发帖数: 1187 | 8 did u speed vac? completely dry pellets are hard to dissolve. did you get
very high conc. in your solution? (>1ug/ul) |
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B*******d
发帖数: 34 | 9 Please try real time PCR and bioanalyzer,
No single method is perfect. all have different degrees of CV.
Nanodrop is not accurate when the conc. is lower than 5ng/ul. |
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g*********r
发帖数: 9366 | 10 confirm that is protein first
then optimize the conditions
change the conc. of everything in the buffer ( salt, buffer, pH, precipitant
etc) |
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w******e
发帖数: 1187 | 11 the thing is, I'm not measuring the value of the target sample,
instead, I have a fix protein conc. for my target sample, 0 protein
for my control. My purpose is instead to demonstrate that my assay
indeed worked in detecting the protein. hence the headache...
maybe I worried too much. all I need to show is that the assay
worked, anyway.
compare all the data together. |
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d**********2
发帖数: 103 | 12 Thanks for the information:) And I will try to make things formal:)
To be clear, I was looking at the proteins that could activate the caspase 3
under transfection condition, but not transfecting cells with caspase 3.
Also, I tried different time/conc of STS treatment...did not work...i could
alway only see the full length caspase 3. So if you dont mind can you tell
me what antibody you use? Like company and catalog num? Thanks alot and I
appreciate it:) |
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T**********t
发帖数: 1604 | 13 I would try again with lower IPTG conc. (say, 0.4mM) and lower temperature (
say, 33C).
Also, if your protein is toxic to E.coli cells, it helps if you use the
strain BL21(DE3)pLysS. |
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g*********r
发帖数: 9366 | 14 yes, try low IPTG conc. even down to 0.1 mM
and try room temp inducing and growing aftrwards
( |
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j********r
发帖数: 156 | 15 As to 3MA, it is hard to dissolve it. The following is the method I always
use to prepare 200 mM stock in water:
For the Sigma 3MA (M9281-100MG)
1. Heat a large amount of tap water (say 500-1000 ml) to around 70 celsius
degrees.
2. Heat 3.352 ml tissue culture-grade water in a tube (eg. the tube for
flow cytometry or the regular 15 ml conical tube), by sitting it
in the heated water.
3. Remove the whole stuff into a hood (the temp of the tap water can
sustain for a while w/o a heater... 阅读全帖 |
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n***w
发帖数: 2405 | 16 Thank you guys.
I use PGEX2T vector so GST is on the N-terminus.
I normally do 100ml culture size. Here is how I did this:
1. small culture of the bacteria from glycerol stock O/N, about 6-8ml.
2. transfer the small culture to 100ml 2xYTA medium in the next day morning
, shaking at 300rpm, 37degrees.
3. it ususally takes about 1hr 40min - 2 hrs to have an OD600 around 0.75.
4. add IPTG (stock 100mM) 100ul to get a final concentration of 0.1mM.
5. Lower the temp to 22degrees and shake at 300 rmp... 阅读全帖 |
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o*****r
发帖数: 156 | 17 sometimes for old buffer, the ATP might be hydrolyzed
add additional ATP to 1 mM (final conc) would help for the ligation. |
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x********u
发帖数: 430 | 18 Thanks for your reply.
We use same inducer for all the experiments.
P should have same function with respect to X1-R2 and X2-R2. Once bound with
X1 or X2, both R1 and R2 should block the transcription of eGFP.
R1 and R2 recognize different inducer. The inducer for R2 is a universial
signaling molecule within all living organism.
One model currently I came up with is that X2 will interact with X1 and
decrease the binding affinity between R1 and X1, thus we saw an activation
effect when X1 is comb... 阅读全帖 |
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b**********8
发帖数: 349 | 19 最近开始做这个实验，大概流程如下，RSB+0.5% NP-40 分离nuclei，然后用NEB 公司
的 DNase I （浓度分别为0,8,16,32,64,128 Units/ml）在37度切25min，加EDTA(
final conc 5mM)混合后75度10min终止反应，蛋白酶k 56度过夜消化，酚氯仿抽提。取
2ug 基因组DNA跑0.8% TBE 胶，理论上随着DNase I 酶量加大，应该能看到一个DNA逐
渐变碎的过程，可是我做了几次，都只看到一个很微弱的趋势，我现在有两个问题想请
教大家，
1）是否一定要在DNase I 处理后看到一个明显的基因组DNA变碎的趋势？
2）NEB网站上建议的DNase I处理完毕后需要加一定量的的EDTA，再75度10min，但是我
看别的文献里说EDTA是用来保护mRNA不受降解的，适用于做reverse transcription之
前去处RNA样品中残留的基因组DNA。我这个实验里需呀尽量除去RNA，是否可以不加
EDTA而直接做heat inactivation？
请大家帮助，谢谢 |
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a*****x
发帖数: 901 | 20 1uM is misleading. You don't know the conc. in blood or tissues. You have to
do PK/PD for that. |
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o*****d
发帖数: 11 | 21 10L左右的话，GE wave system比较方便，主要是整个cellbag是disposible的
也可以用stirred bioreactor
你可以做10x的scale up。比如 1L 到10L
CHO-S（invitrogen）的话比较皮实，1E5/mL都没太大问题
293F (invitrogen)starting conc. 3-5E5/mL
media的话，如果你不要求高密度，DMEM/F12, rpmi都可以
如果加血清的话更好
很多公司都卖serum free medium
拿CHO-S 做例子，有invitrogen, hyclone, irvineSci等等
wave cellbag不用灭菌，传统的glass flask用蒸汽就可以了
inoculation的话 上youtube看看教程吧
你们表达啥蛋白啊？ outsource是最好的选择了吧
又快又省心 |
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i***0
发帖数: 160 | 22 To be honest, according to my experiences, the worst reliability of a plate
reader is the absorption assays. I have tried Bradford or OD280, both give
high standard deviation. However, if you just want a guess, try using your
sample at original conc, 1:1 dilution, and 1:4 dilution in triplicate. You'd
better have BSA standards at least 5 concentrations in the same plate in
triplicate. These will give you a relatively reliable readout. OD is linear
at 0.5 to 1, roughly. good luck |
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l***s
发帖数: 841 | 23 That's weird. We have tried thousands of primers from the database, and they
mostly worked well (>90%). As to isoforms, the primers were designed to
cover all known isoforms of a gene based on NCBI RefSeq. This was done by
designing primers from the common regions of all isoforms. Not sure why your
Ct values are low. Did you follow the protocol from the website (primer
conc, annealing temperature, etc)? Another consideration is that not all
genes are expressed in all cells.
WELL |
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j*b
发帖数: 341 | 24 How do u know hyper polarization increases oxidative stress?
What is the mechanism?
For LZ, you can try different conc. oligomycin
induce |
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n**********u
发帖数: 77 | 25 qpcr的eff%太低了，around50%, BSA的final conc是0.8micro gram/micro liter, 是
网上推荐的最大的值了，还有什么原因呢？ 标准曲线最大浓度的一组ct值是4，这个是
不是不太正常？但是sample的ct值都是十几左右，浓度都很大。 |
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w********r
发帖数: 1431 | 26 just re-transform your low conc. miniprep plasmid
the |
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i***l
发帖数: 1656 | 27 i am telling you the trick i used
you are questioning me about my trick
what do you want to hear from me?
before asking"3-4M urea会不会太高了" have you check why use urea and what's
the usual range of urea used in denaturing buffer?
what's the conc. you think is better? |
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g******r
发帖数: 139 | 28 OK,what you mean is the cDNA concentration used should be in the linear
region of the reaction.
This concentration would vary for each of the genes, dependent on the
abundance. I did not perform such cDNA dilution pilot experiment for each
gene. I just used 2-4 ng cDNA in the real-time PCR. One would assume the
gene with Ct of 25-31 is within the linear range, if the Ct is >33, it is
possible the cDNA conc is kind of out the range.
And it is true as Isaid that I got no outlier replicates for gen... 阅读全帖 |
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r******g
发帖数: 600 | 29 在使用各种inhibitor之前要做一个 各种inhibitor concentration的titration assay
你知道吧？
manual里面有
你要optimize你实验各种inhibitor的conc |
|
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p**********m
发帖数: 472 | 31 collision theory 说分子只有碰撞才会发生反应，这很好说明了为何reaction rate
depends on concentration.
但是一级反应根本没有碰撞，可是这个反应速度跟conc.却是大有关系
这个矛盾大家如何解释呢？ |
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l**j
发帖数: 651 | 32 有可能是prep 21 mm 挺常用的
prep进样通常在1ml crude conc. ~ 20 mg/ml
不过 就算是prep 楼主那个也overload了 5 倍 |
|
v*****e
发帖数: 134 | 33 This is actually not easy as it might sound.
Assuming dilute solutions. With all pka, pkb known, it is mathematically
possible to solve all equations together. But in reality, these equations
are often of high degree and with multiple variables(多元高次方程). A
practical way to solve it is to use iterations (牛顿迭代法）.
An even more practical solution is to measure the pH and calculate every ion
conc accordingly, as 楼上 said. |
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p******1
发帖数: 1388 | 34 what's relationship between pka of analyte and PH of mobile phase? if you
need to assay a basic organic compound.
how to make 1000ml buffer in water with PH=2, 4, 6, 10....? if i give you
pka and conc., and a calculator.. (do not use google.com)
What's meaning of validation? how to do a validation ( do you know how to
write an impurity method validation protocol? if i give you Label claim, and
impurity limit (%), plus some general "FDA laws"...)
what's main differences between assay, impurity, c... 阅读全帖 |
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r*****0
发帖数: 366 | 35 用的是mass hunter 一些参数不明白，也没人可问！
谢谢！
建立calibrations curve,用浓度和ratio,那个ratio是什么？
在elementsal report 里的conc. 是怎么来的？和cal curve 里，浓度是什么关系？ |
|
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c*******n
发帖数: 1648 | 37 I am a layman of bio stuffs, but how about phase separation method? For
example, say you got a median quality solvent for PLGA and their phase
diagram. You may manipulate conc.,temp, cosolvent, and other parameters of
kinetics of phase separation to get particle size that you want! |
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b******0
发帖数: 45 | 38 1. purify styrene by basic alumina column
2. conc. is too low. try polymerization without toluene if A can be
dissolved in styrene. Or just add 0.25~0.5ml toluene if the solubility of A
is not poor
3. three days seems too long using AIBN as initiator at 80C |
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w********b
发帖数: 356 | 39 Both endothelium and smooth muscle have receptors for Ach. The endothelium
receptors cause NO,PGI2 and EDHF production, producing endothelium-mediated
dilation. The smooth muscle receptors, however, cause contraction. At
lower concentrations of Ach, the endothelium receptors are activated and
produce dilation. At higher conc of Ach, the smooth muscle receptors are
activated and produce constriction. |
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y****z
发帖数: 27 | 40 Lao Dao , you make people warm and encouraged. Thanks
Conc for a step-up on your career. |
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k**e
发帖数: 2728 | 41 ☆─────────────────────────────────────☆
ctsm (sharpshooter) 于 (Sat Sep 25 10:58:10 2010, 美东) 提到:
发信人: USMedEdu (US_CMGs), 信区: Tennis
标 题: 要去加拿大了，酸一把。。。。。。
发信站: BBS 未名空间站 (Thu Sep 23 18:06:48 2010, 美东)
要去加拿大了，酸一把。。。。。。
20年前洋插队第一站是加拿大落的脚，后来到美国讨生活。转了一大圈，又要回加
拿大讨饭了，酸一把。。。。。。
好在社会主义加拿大生活舒适安逸铁饭碗，俺决定以后每年回NY看USO顺便参加
球友爬梯，会会老朋友。
六言．有感
力刀
来时枫国草绿，归去叶将枯黄。
当年理想成灰，怒视坦克机枪。
北极凌厉朔风，风城刺骨雪霜。
来时孑然一身，患难夫妻儿郎。
十年坎坷漂泊，铁心重上考场。
一分过线分数，不惑住院奔忙。
离家两载三处，专科又是一双。
募捐艾滋孤儿，海外国内同行。
码文辅导学友，血泪熬墨文章。
学成已知天命，鬓霜少年心狂。
钱财名利如烟，输赢又有何妨... 阅读全帖 |
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h**l
发帖数: 4883 | 42 不用全背。 几个重要的要知道。
比如loading dose=Vd*target conc
CL=Vd*Ke
t1/2=0.693/Ke
maintenance dosing rate=CL*Css
F=AUCoral/Dose oral/AUCiv/Dose iv
Cockcroft-Gault formula |
|
b***k
发帖数: 2673 | 43 ☆─────────────────────────────────────☆
haotianqi2 (jacky) 于 (Sat Oct 6 01:37:28 2007) 提到:
Some questions listed below from this big-name IB quant exams given this
week. As usual there are four sections(basic maths, advanced maths and
brainteaser,mathematical finance and CS). Unlike the one given in January,
my feeling is that undergraduate maths were stressed(Series, Linear Algebra,
Geometry) this time and there are much less brainteasers and probability
questions; for CS part, more conce |
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m*******r
发帖数: 4468 | 44 第二次， 从混了酒精的水瓶子里面， 转回来q里面不是带了小部分酒精吗？ 所以酒精
conc. 还是要高吧？ |
|
m******g
发帖数: 91 | 45 To go further, I think if you can do the followiing exp.s:
1) just heat FeSO4, w/o other salines
2) try other source FeSO4 products (not the same company)
3) try other water (ddH2O, and autoclaved)
4) Cool down the sol. and see if the color will come back to green
5) try diff. Conc. FeSO4, diff. Heating Temp., draw a O.D. curve and see if
there is any dose- or temp- dependent relations
etc. and see |
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g*******y
发帖数: 380 | 46 仔细看了一下自己的code，感觉明白你说的意思了。
问题出在avg(a.conc)上面是吗？这个a还是在用原来的数据，而不是根据criteria
match之后剩下的数据，要实现我的想法，这个a要在下面的select里面写一个view让他
们先subset才行。
谢谢。
ave
agai |
|
l******o
发帖数: 3764 | 47 怎么能弄出个legend来，显示红线和蓝线都是什么？
查了半天也没查到
my code
====================
pl <- ggplot (tab.dat) +
geom_point(data=tab.dat, aes(TAD, DV), colour="#444444", size=3, shape=1
) + #default shape is closed circle
geom_line(data=vpc.dat, aes( (vpc.dat$upper+vpc.dat$lower)/2, vpc.dat[["
X50.sim"]]), size=1.5, colour="#882222") +
geom_line(data=vpc.dat, aes( (vpc.dat$upper+vpc.dat$lower)/2, vpc.dat[["
X5.sim"]]), size=1.5, linetype="dashed", colour="#222288") +
geom_line(data=vpc.dat, aes( (vpc.d... 阅读全帖 |
|
s**r
发帖数: 669 | 48 paired Wilconxon test其实和paired T test的null hypothesis都是一样的，就是H0:
mean(Read)=mean(Conc),从data的range来看p-value很小是很正常的。 |
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a*******g
发帖数: 80 | 49 刚看见你的read 和 conc 差这么大 那当然p很小了 因为paired test 是test paird的
差值是不是0 你这个不可能是0啊 |
|
a*******g
发帖数: 80 | 50 哦 你把乘完以后的conc和read 相减 画个histogram 或box plot 看看location在不在
0 |
|
