h**********r
发帖数: 671 | 1 同上另外一种方法。
Yeast “smash-and-grab” DNA prep:
• pipette 1-2 ml YPD onto transformation plate; scrape colonies off
with the end of a glass slide and pipette into a microfuge tube
• spin down 15 sec; pipette off sup; if cell pellet is more than 50-75
μl, remove excess and discard
• to cell pellet add 0.2 ml lysis buffer, 0.2 ml phenol/CHCl3 and 0.3
g 0.45-0.5 mm glass beads* (I use calibrated scoop made from cut microfuge
tube pierced with syringe needle) – seal carefully because be... 阅读全帖 |
|
h******y
发帖数: 351 | 2 这几种脂肪酸中，Palmitic Acid 和 Stearic Acid常温下是固体，计算摩尔浓度比较
容易。其他几种常温下是液体，但是大体上他们的密度和水的密度很相近，1ml近似1g
。这些脂肪酸都极易溶于乙醇。溶解Palmitic Acid需要略微加热（50C)。
The free fatty acids are toxic unless they are presented bound to purified,
fatty acid free, endotoxin-free serum albumin, which is used at a typical
concentration of 0.1–0.2%.
Take 31ul 1M palmitic acid, 2.8ul 1M Palmitoleic acid etc and bring the
final volume to 1000ul with 100% EtOH, you will get 0.1M FFA. Add 76ul of
the mixture to 1L culture medium and you w... 阅读全帖 |
|
m**********2
发帖数: 57 | 3 我用humerus section做in situ hybridization的时候,用 RNA hybridization
incubation 过夜70度,发现tissue结构收缩,如附件所示,图示是我已经完成整个in
situ的结果,事实上我在incubation以后就已经发现tissue结构收缩了.
我问了一些人说需要在fixation的时候做decalcification,我做了decalcification,结
果in situ 的时候tissue还是会收缩,事实上看起来更严重.
我不知道这究竟问题出在什么地方,看起来像是tissue attach不牢固,导致容易变形.请
问有什么方法可以使tissue attach牢固么?
我是根据下面的步骤做的:
1. cut the arms and legs
2. 4% PFA (pH 7.1-7.4) overnight at 4C.
3. PBS three times for 5 minutes
4. 14% EDTA for decal 3 days.
5. wash with PBS
6. store in 70% ETO... 阅读全帖 |
|
s******y
发帖数: 28562 | 4 如果你要转的是类似Hela, HEK293T 这些大众化的细胞的话，
我亲手做过，用lipofectamine能转。如果你担心内毒素的话，
在做miniprep 的时候额外多洗两次（一次用buffer PB,
然后70%ETOH 洗的这一步多洗一次）
唯一要注意的是大部分mini-prep 出来的质粒浓度比较低，
如果浓度太低（低于50ng/ul) 的话要浓缩。
和线性，而maxi的去除了内毒素，且 |
|
C*********u
发帖数: 811 | 5 最近抽RNA用nanodrop测浓度，发现260/230ratio很不稳定，有时很好有时很低（～0.5
），而260/280 ratio却一直很好。
我已经用70%的ETOH洗两次了，而且在extraction的时候尽量避免protein和pheno污染
了，用的也是分装的nuclease free的水，实在是百思不得其解。
请问大家低260/230ratio会影响之后的RT和qPCR吗？
谢谢 |
|
x**2
发帖数: 255 | 6 Your compound must be very hydrophobic
You can try MeOH or EtOH, both of them are normally used to dissolve
hydrophobic compounds |
|
C*******e
发帖数: 4348 | 7 “但是拿到基因组DNA上扩增就很差。 不同的底物浓度的ct值
很接近，而且都偏大”
——除了上面这么多id说的，
另外还有一个情况要注意，gDNA提取过程中很多时候会带有PCR inhibitor
有的是样品带来的（比如environmental sample可能含humic acids， polyphenols
），有的是提纯过程中引入的（比如EtOH去的不干净之类的）
所以不是模板越多就越该P出来的
很多时候稀释模板以后反而能够P出来就是这个道理
因为稀释的时候PCR inhibitor也被稀释了
你这个“不同第五浓度Ct值很接近，而且都偏大”可能就是这个因素 |
|
S******9
发帖数: 2837 | 8 可以用。
我现在打新生鼠还不能用etoh配，单纯的oil配，需要70-80度溶解药物，然后分装，每
次用以前只能再次65度溶解以后快速赶到老鼠房去打针。 |
|
|
l**********1
发帖数: 5204 | 10 which holds lower density? water or EtOH, lx ID s/he forgot that.... |
|
S******9
发帖数: 2837 | 11 第一次做MS model，临床评分不错，Luxol fast blue-HE染色总是不理想。
slides是frozen slides， 开始脱水到95% etoh，然后开始60度过夜Luxol fast blue
溶液，次日进行后续步骤。
感觉蓝色和红色对比不鲜明。
请有经验的指点一下。
谢谢 |
|
b*****s
发帖数: 169 | 12 做酶切鉴定或是测序，我从来不用kit，效果是good enough，已经测序有近2000个样品
了。只有做转染才用kit。
我的方法是挑菌落到1ml LB in 1.5ml eppendorf tube, culture for 16h, 吸取50-
100ul culture in 96-well plate 放在4oC 用于以后正确克隆的扩大培养。离心
13200rpm for
0.5min. The pellet is resuspended in 200ul P1, then P2, then P3, spin at
13200rpm for 15min after stored at 4oC for 30min. add equal volume
isopropanol to the supernate in a new tube, spin for 15min, wash with 70%
ETOH. 然后溶于50-100ul水或是TE即可，浓度在150ng/ul 左右。一天做96个克隆是不
难的。
这种样品测序时我试过两个公司，同一样品，genewiz可以得到23个有用的... 阅读全帖 |
|
n***w
发帖数: 2405 | 13 add a little bit EtOH and vortex extensively and it should turn clear. |
|
r***n
发帖数: 15 | 14 想用一端是SH另一端是NH2的ligand来合成AuNP，S连在Au上，NH2暴露在外面做后面的
反应。合成方法选用在EtOH里用NaBH4还原HAuCl4。但是alkyl amine本身也可以用来稳
定AuNP，虽然说SH应该是比NH2更强的配体，但是还是担心NH2的存在会不会对AuNP的合
成造成影响。
另外，SH和NH2是处于酸碱平衡中，大部分的存在形式是S(-)和NH3(+)。不知道这个会
不会影响S和Au第一步反应的前驱物的形成。
有人做过类似的合成吗？多谢指点。实在是这个ligand有点贵，100mg要300刀，不太敢
乱试。要尽量多的将NH2连上去，所以暂时不考虑exchange的办法。 |
|
w*****g
发帖数: 317 | 15 EtOAc+MeOH or EtOAc+EtOH我都用过，都很好。 |
|
b**s
发帖数: 589 | 16 what 沉淀剂 have you tried, what about EtOH? you might try neutralize it
yes, 透析膜 is a good ideal
我合成的高聚物,主链是CH2CHO,在CH上连着侧链CH2P=O(OEt)2,经水化得到CH2P=O(OH)2,
DMAC
它?
报了 |
|
d****n
发帖数: 237 | 17 我使用到一种甲基丙烯酸酯单体，(N,N-dimethylamino)ethyl methacrylate,
DMAEMA, 已经将它聚合成PDMAEMA，现在有什么办法能把它水解掉或者醇解掉？用一般
的碱催化水解似乎没有效果，尝试过H2O/ETOH的醇解，文献中说，对于DMAEMA单体有效
，但是对聚合物几乎没有效果。诸位还有别的办法吗？
查查了其它文献，常见的是利用碳酸酯而非羧酸酯的水解，甲基丙烯酸酯中水解多是
叔丁酯。
thanks and bow. |
|
d******a
发帖数: 127 | 18 >> 准备戒网了！！
Why?
戒网 is much more difficult than 戒 smoking/EtOH. |
|
N****e
发帖数: 31 | 19 38yr, female, present to PAH on 4/4/10 with diarrhea for 4 days, initially
loose watery diarrhea, 3-4 times/days, now worse and loose watery stool
every hour, and blood mixed with the stool, medium volume, non-foul smelling
, with mild crampy abd pain. Have had 5 BMs since yesterday, and the last
one almost all blood. (+) subjective fever, no chills, no cough/SOB. Had a
course of antibiotics (Keflex) 2 weeks ago (3/9/10- 3/14/10).
PMHx: Kidney stones, Seizure disorders, Vit D deficiency
PSHx: S... 阅读全帖 |
|
A*******s
发帖数: 9638 | 20 This case is tough. 20 WB for the first. (from BZ)
A 35 yrs old WF, was sent to ER by her husband in AM since he could not wake
her up.
The patient was entirely normal before last dinner. They planned to have
BBQ, but after she drank a can of beer and 3 cans of soda, she felt sick. So
she went to bed directly.
At ER, she was stuporous, a drug screen was neg. CT of brain, EKG, CXR were
all normal. LP is neg. UA(-), Blood culture was sent. All lab was normal
except Glucose was 60. So she was gi... 阅读全帖 |
|
A*******s
发帖数: 9638 | 21 The fact was that she got worse after she received D5 1/2NS.
And I repeat: Drug screen is neg for ETOH. So no intoxication.
hypoglycemic |
|
y******a
发帖数: 590 | 22 interesting case. I thought EtOH intoxication can suppress the resp. center
, cause hypoxia, then bradycardia. metabolic acidosis can cause bradycardia
and hypotension too. Never thought about vitamin deficiency. Thanks.
But
giving |
|
A*******s
发帖数: 9638 | 23 I think you know the answer, don't you?
This has something to do with ETOH metablism.... Anybody wants to elaborate
? |
|
A*******s
发帖数: 9638 | 24 我被问过两个问题， 1. What do you think about VA system?
2. What is your opinion on ETOH abuse? |
|
A*******s
发帖数: 9638 | 25 Do you mean ＂physicians with interest on ETOH abuse"?
I guess all Chairs /PDs are some kind of politicians. So they do care about
social problems very much. However, they do not count on your opinions.
They want to see your insight, your maturity and your logics.
It does not matter what your PD's interest is.
|
|
p********g
发帖数: 372 | 26 来自主题: MedicalCareer版 - 啥么意思？ Trauma Panel = CBC, chem 7, Coags, Type and Cross, amylase, ABG, ETOH level,
U dip and Utox, (hCG for women of child-bearing age) |
|
A*******s
发帖数: 9638 | 27 昨天有同仁对我的一个case感兴趣， 我今天查了下病历， 给大家写个完整的。
53 yrs old WM, previously healthy, presented with intractable fatigue and
weight loss for 3 months. A comprehensive workup by his PCP including a CT
of chest was reported as unremarkable.
PMH: Asthma, seeing a pulmonologist who reviewed CT and agreed with
radiologist's report.
Soical. Smoker 1 ppd. No ETOH/Drug.
FMH: neg for CA.
ROS: Not remarkable, no diplopia, dysphagia, no fever, no joint pain, no
muscle ache.
PE: Vital normal.
No positive findings on P... 阅读全帖 |
|
s********o
发帖数: 3319 | 28 C?
EtOH --> chronic pancreatitis --> decreased HCO3- secretion |
|
z*****g
发帖数: 558 | 29 Today, the attending talked about how to do a case presentation. Here is a
summary about his talk:
1: HPI is about 70% of the whole case. Do present the admission time and
reasons. Do mention if the patient looks sick, toxic or fine
2: PE should be less than 10%, which includes mainly positive signs. For
other negative systems, we should describe them in a little bit detail
rather than just saying within normal limit, such as “could not feel
spleen or liver”
3: Review of systems is the least th... 阅读全帖 |
|
o******u
发帖数: 60 | 30 想說試試看...
看有沒有人感興趣...
主要是挑書中錯誤以及自己經驗反饋
分享知識
原則一星期一次
時間待定
如有興趣請跟貼回復並附上自己的Email address,
或是可以消息我您的email address.
Request
1, at least pass step 1, 2, score for step 1,2 both above 95.
2, at least have 3 self critics or comments in every page and they are not
written in the book.
3, have bought the book.
4, can link the knowledge through from step 1 toward step 3.
for example, Page 75,
Porphria Cutanea Tarda
my comments:
1, this is the disease called Vampire disease, you afraid light, you pull
out se... 阅读全帖 |
|
m******r
发帖数: 1904 | 31 1.没用过。不懂。
2.Common abbreviations for SP note
BP Blood pressure
P pulse
R respiatory rate
T temperature
yo year-old
m male
f female
b black
w white
L left
R right
hx history
h/o history of
c/o complaining of
NL normal limits
WNL within normal limits
Ø without or no
+ positive
- negative
Abd abdomen
AIDS acquired immune deficiency... 阅读全帖 |
|
m******r
发帖数: 1904 | 32 ☆─────────────────────────────────────☆
oldclock (oldclock) 于 (Wed Apr 18 10:59:11 2012, 美东) 提到:
1。step2 CS的UW video要单买还是在CS的UW题库里都包括了？
2。patient note可以写ABBREVIATIONS吗？除了FA上的ABBREVIATIONS，还有其他哪些
地方的ABBREVIATIONS可以用呢？
3。此外，看到麦地里有提到过patient note用子弹体方式书写，什莫是子弹体啊？哪
里可以找到范文呢？ 给扫扫盲吧。
多谢各位了！
☆─────────────────────────────────────☆
medwater (小麦) 于 (Wed Apr 18 11:08:03 2012, 美东) 提到:
1.没用过。不懂。
2.Common abbreviations for SP note
BP Blood pressure
P pulse
R respiatory rat... 阅读全帖 |
|
d*********n
发帖数: 18 | 33 this one of my CS protocols, hope can help whoever going to take the test, I
don’t need any more.
not perfect, but good to figure out diagnosis, should not miss the diagnosis
,
do not need to ask all questions , must adjust base the case,
if better ideas, pleas modify to make easier for people who may need
Amenorrhea
Mental check list:
Pregnancy,
hyperprolatinemia,
hypothyroidism,
PCOS,
premature menopause,
depression
asheman syndrome,
anorexia nervosa,
strenuous exercise
Patricia Garrison, a 36... 阅读全帖 |
|
d*********n
发帖数: 18 | 34 Confusion case: O CFDP P ADL, IADL, TIA/stroke, depression, DM, thyroid,
normal pressure hydrocephalus, PAMHUGSFOSS
insulin induced hypoglycemia
Electrolyte abnormalities
Vitamin B12 deficiency
Medications
TIA
Multi-infarct dementia
Chronic subdural hematoma
Alzheimer’s dementia
Normal pressure hydrocephalus
Intracranial tumor
Neurosyphilis
Depression
Hypyothyriodism
DDx Confusion
DEMENTIA
D-Diabetes /Dementia/ Drugs
E-Epilepsy
M-Migraine/Mult Infarct Dementia
E-Ethanol (withdrawl / Toxicity)
N... 阅读全帖 |
|
A*******s
发帖数: 9638 | 35 你说的都是对的，但你看清这个题的问题：the most appropriate Initial ....
相信一个有ETOH问题的病人你会后悔的，信不信由你。：）
patient
and
physician
a |
|
a*****s
发帖数: 3643 | 36 coat gold or surfactant or metal oxide
Store in EtOH |
|
w********g
发帖数: 186 | 37 less EtOH or use aceton which will evaporize more rapidly |
|
r***n
发帖数: 15 | 38 想用一端是SH另一端是NH2的ligand来合成AuNP，S连在Au上，NH2暴露在外面做后面的
反应。合成方法选用在EtOH里用NaBH4还原HAuCl4。但是alkyl amine本身也可以用来稳
定AuNP，虽然说SH应该是比NH2更强的配体，但是还是担心NH2的存在会不会对AuNP的合
成造成影响。
另外，SH和NH2是处于酸碱平衡中，大部分的存在形式是S(-)和NH3(+)。不知道这个会
不会影响S和Au第一步反应的前驱物的形成。
有人做过类似的合成吗？多谢指点。实在是这个ligand有点贵，100mg要300刀，不太敢
乱试。要尽量多的将NH2连上去，所以暂时不考虑exchange的办法。 |
|
b******n
发帖数: 80 | 39 请教高手：
为了产生H2Se，我用Se粉分散在EtOH中，然后用NaBH4水溶液还原Se到NaHSe，整个过程
用N2保护，然后用另外一个管道通入Cd（NO3）2的水溶液中，用sodium citrate作为表
面活性剂。
当Se充分溶解后，希硫酸溶液加入前一个反应瓶，可以看到剧烈的气体产生，这时由缓
慢的N2流入第一个反应瓶来推动H2Se气体到第二个反应瓶去，反应了半天，一点颜色变
化都没有，说明根本就没有和Cd2+反应。
这时，我加大N2的流量，发现第二个反应瓶迅速变成红色，同时收集废弃的瓶子也变成
红色，这应该不是H2Se。过了一天，红色溶液底部有褐色沉淀物。
问，到底是SeO2还是Se？
我整个过程有哪里出错，为什么没有CdSe NPs？
BTW，
1. H2Se极错，我用防毒面具也能闻到，什么办法可以隔绝这玩意？
2. 我做了阵子CdS和CdSe，回家居然闻不到茉莉花的香味，我爱人和朋友说那个香味非
常浓，我怎么没闻到，我是不是杯具了？ |
|
y***e
发帖数: 6082 | 40 你是照谁的文献做啊，彭晓钢或者Paul的文献比较有权威的说，我当年是照着彭的那篇
JACS做的
请教高手：
为了产生H2Se，我用Se粉分散在EtOH中，然后用NaBH4水溶液还原Se到NaHSe，整个过程
用N2保护，然后用另外一个管道通入Cd（NO3）2的水溶液中，用sodium citrate作为表
面活性剂。
当Se充分溶解后，希硫酸溶液加入前一个反应瓶，可以看到剧烈的气体产生，这时由缓
慢的N2流入第一个反应瓶来推动H2Se气体到第二个反应瓶去，反应了半天，一点颜色变
化都没有，说明根本就没有和Cd2+反应。
这时，我加大N2的流量，发现第二个反应瓶迅速变成红色，同时收集废弃的瓶子也变成
红色，这应该不是H2Se。过了一天，红色溶液底部有褐色沉淀物。
问，到底是SeO2还是Se？
我整个过程有哪里出错，为什么没有CdSe NPs？
BTW，
1. H2Se极错，我用防毒面具也能闻到，什么办法可以隔绝这玩意？
2. 我做了阵子CdS和CdSe，回家居然闻不到茉莉花的香味，我爱人和朋友说那个香味非
常浓，我怎么没闻到，我是不是杯具了？ |
|
o****n
发帖数: 9475 | 41 Adm via ED 2 days ago with blood glucose 700. Dx hyperplycemic Hyperosmolar
Nonketotic Syndrome
Medical and surgical history
Type 2 Diabetes x 4 yrs.
Hypertension (HTN) x 8yrs
Coronary artery disease (CAD) x___yrs
Alcohol (ETOH) abuse x 40 yrs
L inguinal herniorrhaphy x 10yrs
Cholestectomy x 15 yrs |
|
M******C
发帖数: 623 | 42 ☆─────────────────────────────────────☆
oxhorn (^_^) 于 (Mon Mar 21 20:55:02 2011, 美东) 提到:
type 2 病人急诊到ED，诊断了HHNS，hyperglycemia hyperosmolar Nonketotic state
,两天后Head to toe assessment:
IBW 150%
I/O 3600/1600
+2 edema L ankle
pedal pulses +0/3 bilateral
lung crackles
last BM 4 days ago
FSBS 200
ate 100% of lunch，complain of still hungry
HHNS的症状不是dehydration吗？什么情况下会I/O +？priority nursing diagnosis可
以是Excess fluid volume吗？
Excess fluid volume r/t decrease fluid volume output AMB I/O(+ 2000 ml)... 阅读全帖 |
|
A*******s
发帖数: 9638 | 43 昨天有同仁对我的一个case感兴趣， 我今天查了下病历， 给大家写个完整的。
53 yrs old WM, previously healthy, presented with intractable fatigue and
weight loss for 3 months. A comprehensive workup by his PCP including a CT
of chest was reported as unremarkable.
PMH: Asthma, seeing a pulmonologist who reviewed chest CT and agreed with
radiologist's report.
Soical. Smoker 1 ppd. No ETOH/Drug.
FMH: neg for CA.
ROS: Not remarkable, no diplopia, dysphagia, no fever, no joint pain, no
muscle ache.
PE: Vital normal.
No positive finding... 阅读全帖 |
|
A*******s
发帖数: 9638 | 44 昨天有同仁对我的一个case感兴趣， 我今天查了下病历， 给大家写个完整的。
53 yrs old WM, previously healthy, presented with intractable fatigue and
weight loss for 3 months. A comprehensive workup by his PCP including a CT
of chest was reported as unremarkable.
PMH: Asthma, seeing a pulmonologist who reviewed chest CT and agreed with
radiologist's report.
Soical. Smoker 1 ppd. No ETOH/Drug.
FMH: neg for CA.
ROS: Not remarkable, no diplopia, dysphagia, no fever, no joint pain, no
muscle ache.
PE: Vital normal.
No positive finding... 阅读全帖 |
|
y***d
发帖数: 33 | 45 在板上潜水有一会了，好像发case的不多啊。是不是这儿attending太多，都没时间写
case了呢？我来抛砖引玉好了。
从今天开始，每星期我会上一个case。 周一history, 周二physical， 周三initial
labs/work up, 周四additional work-up, 周五final diagnosis. 欢迎大家跟贴讨论
。这些case基本上是面向内科resident的。不过attending们也可以复习一下哈。如果
受欢迎的话我会一直贴下去。废话到此为止。 先上第一个case试试反响如何。
CC: bilateral leg swelling and diarrhea
HPI: 72 yo f h/o HTN, diarrhea and pneumatosis intestinalis of unclear
etiology presented to ER for worsening bilateral lower extremity edema and
ongoing diarrhea.
Initially presented 2 month PT... 阅读全帖 |
|
y***d
发帖数: 33 | 46 Looks like people enjoyed last week's case. Here's the second one. This one
is shorter and more straightforward.
CC: fever, abdominal pain and arthralgias
HPI: 31 yo merchant ship worker p/w 3 days of acute febrile illness. 16 days
PTA, he flew from India to Long beach, where he had a negative pre-ship
exam including labs. Then patient boarded an oil tanker and sailed up west
coast. 3 days PTA, he developed mild mid-epigastric and LUQ pain. Over the
first day, pain worsened and was associated wi... 阅读全帖 |
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n****a
发帖数: 2525 | 47 Thanks everyone for replying.
It's been 12 days into her hospital stay, she is still in pain. Her IgG4 to
look for autoimmune pancreatitis was negative, lab ordered for possible
Celiac (which leads to poor absorption of Levothyroxine and pancreatitis
link) also negative. GI still thinks pancreatitis, might have been
exacerbated by cold meds she was taking that has some ETOH content. EGD was
done by GI only showing gastritis. CT ab showed nothing but cysts, no
pancreatic cyst or pseudocyst or oth... 阅读全帖 |
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A*******s
发帖数: 9638 | 48 PMH: DM, type 2.
Social: No Smoking/ETOH/Drug abuse.
FMH: No history of movement disorder.
ROS：Not remarkable.
PE: VSS: Normal
Except chorea movement and areflexia, all other exams are normal. |
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A*******s
发帖数: 9638 | 49 VA is great for someone who just wants an easy life. But once you have
worked at VA, you probably would change your mind.
I assume you are a resident, so my advise is not to consider it at all. It
is not good for your career development and once you are in, you will be
stuck in that system.
Benefits are great but you can make much more than that. A good example is:
I used to work for a hospital with great benefits. I even had a pension plan
. After 4 years, I left for a clinic practice. For a s... 阅读全帖 |
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A*******s
发帖数: 9638 | 50 Defensive medicine.
Like kobejordan's case, ETOH toxicity caused some neurological deficits, we
have to run a stroke workup. Medicare will pay over $10,000 to the hospital.
How much should cost? A fee for EMT, a good nose from one EMT guy and a
drug screening. At most $1000.
should |
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