s****9
发帖数: 932 | 1 No perfect methods. It largely depends on your research purpose.
Cells carrying beads are bad for their trafficking in vivo. But magnetic beads enrichment is quick and relatively economic, as compared to sorting. For in vitro study like RNA purification or culture, beads purification is fine.
Sorting is expensive and labor-intensive (I suppose your lab do not have
your own sorter and do not sort cell frequently). But without beads binding
, cells are more healthy. |
|
h**********r
发帖数: 671 | 2 以前做Neurospora的knockout的时候，看到过yeast方面的protocol。感觉他们挺严谨
的。但我不保证work。摘自：
http://www.dartmouth.edu/~neurosporagenome/Projects_files/Proje
Prepare yeast DNA with the Gentra yeast DNA kit:
(the following is slightly modified from their protocol:
http://www.gentra.com/pdf/01160.pdf)
the solutions are available from Gentra as Puregene kit D-6000A or
separately; if you have their Puregene solutions for Neurospora DNA preps,
you only need to order the Cell Suspension Solution (D-6001) and Lytic
Enzyme Sol... 阅读全帖 |
|
p*******r
发帖数: 59 | 3 Career Opportunities at JCVI Infectious Disease (8 Positions)
The PIs did not ask me to post here, I did this just in case someone is
looking for similar positions but did not see these, and I would be happy to
see some chinese fellows to come. All the PIs are very nice.
All listed at
https://careers.jcvi.org/careers/Careers.aspx
http://www.jcvi.org/
No. 1 & No. 2 Post-Doctoral Fellows
Job Responsibilities
JCVI is looking for two Post-Doctoral Fellows to join the Infectious Disease
Group in our ... 阅读全帖 |
|
b********n
发帖数: 22 | 4 Postdoctoral Fellowship – Restriction Enzyme Division – Job Code 1042HZ
The Restriction Enzyme Division invites applications for the position of
postdoctoral fellow. Projects will be part of a basic research program
focusing on restriction enzymes specific to 5-hydroxymethylcytosine. The
project will involve cloning, overexpression, purification, assay-
development, and engineering of such enzymes, and their application to
questions of epigenetics. Research will be undertaken in a highly
coll... 阅读全帖 |
|
c******g
发帖数: 49 | 5
points
also
column purification会把几十bp的primers一起纯化出来，尽管有时候gel上就只看到
一条带，所以我一般都是gel purification。 |
|
y*****3
发帖数: 961 | 6 Sino-American Biotech: Multiple Managerial Bioprocess Position Openings in
China for a US-China joint venture company.
The client company is an existing company well established and joint-venture
with a US Biotech company about over 10 years ago. They were very
successful in supplying Chinese market for diagnostic and reagents in the
past years. Recently they decide to expend the business into biological
product R&D and commercial manufacturing in China for both Chinese market
and global market ... 阅读全帖 |
|
|
l**********1
发帖数: 5204 | 8 问句楼上: 用的是QIAGEN 纯化kit? QIAquick PCR Purification Kit or
Small-Scale Plasmid DNA Purification Kits etc... |
|
l******o
发帖数: 30 | 9 a faculty position in a structural biology lab does not require experience
in protein crystallography? Only "Experiences in molecular sub-cloning,
protein purification protein interaction analysis are required"???? What
kind of "faculty" position is this? A technician?
The
purification
self
relocation |
|
j****t
发帖数: 1663 | 10 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 |
|
l**********1
发帖数: 5204 | 11 alternatively you can try Oligotex-dt30 mRNA purification kit for total RNA
sample:
Manual book:
//catalog.takara-bio.co.jp/PDFFiles/NTrap%20mRNA_j.pdf
Vender link:
//www.mn-net.com/Products/DNAandRNApurification/RNA/NucleoTrapmRNA/tabid/1328/language/en-
US/Default.aspx
Reference:
please go to
PubMed link:
//www.ncbi.nlm.nih.gov/pubmed/18678863
T Yabu - 2008 JBC
31 Oct 2008 – In zebrafish embryonic cells, the endogenous SMase enzyme was
.... using the Oligotex-dt30 mRNA purification kit (Tak... 阅读全帖 |
|
s*******s
发帖数: 623 | 12 Job ID: GF2012A001
Posting Title: Research Scientist/Research Associate/Electrobiochemist
Brief Introduction of Gate Fuels Inc.:
“Imagination, Innovation, Implementation --- Enabled Technologies for a
Green Clean World”
Gate Fuels Inc. is a research start-up firm based in Blacksburg, VA, with
the goal of developing microbial and cell-free platform technologies that
enable low-cost production of biochemicals, biofuels and electricity. The
first platform is based on industrially-safe Bacillus subt... 阅读全帖 |
|
b******r
发帖数: 111 | 13 Sorry for this confusion.
It is too early for me to talk about transgenic mice. I am just during
electroporation of construct into ES cells. My Southern doesn't work for
genotyping. I have to use PCR to isolate positive ES cell clones. Then I
need get a primer pair for genotyping. Forward primer is from Neo, reverse
is from outside. What I did is knockin,not knockout.
Without purification of genomic DNA(just heat to kill proteinase K),PCR
doesn't work. After purification, still doesn't work.
"你... 阅读全帖 |
|
d******n
发帖数: 29 | 14 1.
Towards large scale methods for the selective release of periplasmic human
cystatin C from E. coli
Biotechnology Techniques, Volume 9, Number 3 (1995), 179-184
2.
Optimization of an osmotic shock procedure for isolation of a protein
product expressed in E. coli.
Biotechnol Prog. 2003 Sep-Oct;19(5):1541-6.
3.
Optimization of BLyS production and purification from Escherichia coli.
Protein Expr Purif 39:237-246.
4.
High-level accumulation of a recombinant antibody fragment in the periplasm
of Es... 阅读全帖 |
|
d******n
发帖数: 29 | 15 1.
Towards large scale methods for the selective release of periplasmic human
cystatin C from E. coli
Biotechnology Techniques, Volume 9, Number 3 (1995), 179-184
2.
Optimization of an osmotic shock procedure for isolation of a protein
product expressed in E. coli.
Biotechnol Prog. 2003 Sep-Oct;19(5):1541-6.
3.
Optimization of BLyS production and purification from Escherichia coli.
Protein Expr Purif 39:237-246.
4.
High-level accumulation of a recombinant antibody fragment in the periplasm
of Es... 阅读全帖 |
|
S*********s
发帖数: 304 | 16 // non-denature Tandem purification
3XFLAG + HA
or
ProteinA beads(IgG domain)+HA/FLAG
His or myc affinity and specific 不够~
//denature purification
His and biotag 会比较好~ |
|
d**l
发帖数: 1546 | 17 先把条带切出来， 然后用gel purification kit纯化， 然后再用普通的pcr
purification kit纯化， 然后再做第二轮pcr
一定要两轮纯化 |
|
m***r
发帖数: 49 | 18 帮朋友转发，请直接联系文中的E-mail
One tenure-track assistant professor position is immediately available for
three years in the group of Dr. Gang Cai (University of Science and
Technology of China, Hefei National Laboratory for Physical Sciences at the
Microscale). This is a co-PI position with an initial contract up to three
years and could be extended for a longer time. Successful candidates will
receive a startup package and are expected to perform independent research
in the Cai lab by supervising gradua... 阅读全帖 |
|
|
j**W
发帖数: 89 | 20 PCR 3kb fragment后, 我有用QIAGEN gel purification kit纯化, 再送的测序.QIAGEN
gel purification kit不够好? |
|
q***7
发帖数: 144 | 21 Qiagen做为柱提取DNA的老大，从来没有想着怎么改进他们的产品。他们柱子elute出来
的DNA大都含有洗液，尤其是Gel Extraction, PCR purification， and RNAeasy kits
。你用个连有真空的小枪头在最后一步要elute你的DNA时在柱子里面的那个垫圈上吸吸
看有什么东西。你会看到小枪头里都是洗液。Green BioResearch公司，就想到了这一
点，垫圈上设计了个小洞，这样就不会有洗液留在上面了。这样你elute出来的样品里
就不会含有洗液，而不会抑制后面的酶的活性了。
你可以看一下这个连接
http://www.greenbioresearch.com/gel-extraction-pcr-purification
QIAGEN |
|
|
r******g
发帖数: 600 | 23 最近的nature issue又多了两篇STING papers. 一篇把cyclic GMP-AMP synthase 的
crystal structure resolve了！ 另一片纠正了陈志坚science paper里面的一个小错
误，2‘-5’和3‘-5’ phosphodiester bonds！
仔细看看crystal structure的文章，protein production和purification都是他们自
己做的，不是找陈志坚拿的！ 他们3月21号投的nature，陈志坚的cGAS发现
publication最早在sciencemag online刊登于12年12月底！ 这就意味着，这个
structure组从自己clone，protein production，purification，crystalization，
resolve，writing一共90天完成，太amazing了！ |
|
C***Y
发帖数: 61 | 24 It is necessary to perform PAGE purification for small RNA sequencing
library construction. Based on my experience, Ampure XP beads purification
is not sufficient enough to separate 160 bp from 120 bp. |
|
d******n
发帖数: 29 | 25 J Pharm Sci. 2013 Mar;102(3):813-26.
Investigation of histidine stabilizing effects on LDH during freeze-drying.
Al-Hussein A, Gieseler H.
Pharm Res. 2003 Dec;20(12):1952-60.
Influence of histidine on the stability and physical properties of a fully
human antibody in aqueous and solid forms.
Chen B, Bautista R, Yu K, Zapata GA, Mulkerrin MG, Chamow SM.
Protein Expr Purif. 2011 Nov;80(1):91-6. doi: 10.1016/j.pep.2011.05.014.
Epub 2011 May 26.
Effects of L-arginine on solubilization and purificati... 阅读全帖 |
|
l****5
发帖数: 2968 | 26 请发g***********[email protected]
包子酬谢，多谢大家！
1. Effective natural dye extraction from different plant materials using
ultrasound
2. Anthocyanins: natural colorants with health-promoting properties.
Radish Anthocyanin Extract as a Natural Red Colorant for Maraschino Cherries
3. Separation and recovery of food coloring dyes using aqueous biphasic
extraction chromatographic resins
4. Recovery of anthocyanins from grape pomace extracts (Vitis vinifera L. cv
. Cabernet Mitos) using a polymeric adsorber resin... 阅读全帖 |
|
g*********5
发帖数: 2533 | 27 Isolation of nuclei for the co-IP and whole ChIP protocols is based on the
methods of Gendrel et al. (2002, 2005), Johnson et al. (2002), and Nelson et
al. (2006).
METHOD
For extraction and co-IP of nuclear proteins, see Steps 1-39 (Fig. 1). For
the ChIP procedure, see Steps 40-69 (Fig. 2).
Figure 1.
View larger version:
In this page
In a new window
Figure 1.
Flowchart for the timeline and organization for co-IP of nuclear proteins
from Arabidopsis seedlings.
Figure 2.
View larger versio... 阅读全帖 |
|
c********n
发帖数: 225 | 28 谈谈和蛋白打交道的日子 （七）
引子
光是个奇妙的东西。关于它的来源，各种族的传说都有异曲同工之处 - 中国有盘古开
天辟地带来光明； 圣经创世纪里有很权威的 And God said, Let there be light:
and there was light. 希腊神话普罗米修斯盗火给人类黑暗中带来光明，而自己却被
锁在高加索山的悬崖绝壁上，无怨无悔，忍受着宙斯的惩罚。
这篇咱们来散漫地谈一下生命之光吧， 只是一些体会，不严谨处见谅。
第七篇 生命之光
1.GFP
从一个“天才”对他学生们的challenge开始吧。 - “Tell me， why is GFP so
important to Biology？”
“天才”是他PhD老板在别人面前对他的赞誉。他每天的工作时间在14-16个小时，勤奋
带来的是各种荣誉以及源源不断的基金，即使在grant最黑暗的年代里，他的lab人员也
可以随心所欲的尝试各种方法。他的代价是一个人曲高和寡的寂寞， 在他娴熟悠扬的
小提琴声里慢慢的扩散，扩散。
回到问答， 年代flash back to GFP刚刚投入应用的时候。大家七嘴八舌，它是绿... 阅读全帖 |
|
l****y
发帖数: 398 | 29 我们做了一批anti-GFP和anti-mCherry的nanobody(15 KDa, single-domain antibody)
, 有两个用来做affinity purification/immunocapture很好用。
有需要的同学请通过站内信提供一个US non-profit research institute 的地址.
vector用e.coli 表达，periplasm加Ni-NTA纯化，一般可以拿到2mg/liter culture.
质量比多抗稳定，成本也低。适合需要做很多affinity purification的情况。 |
|
c********n
发帖数: 225 | 30 实验流程第二步：
2. Transfection
更新前
2-1 NLS-NgAgo expressing plasmid is extracted with Wizard® Plus SV
Minipreps DNA Purification System (Promega), and is adjusted to 100 ng/μl
in 0.5x TE buffer (5 mM Tris-HCl, 0.5 mM EDTA，pH 8.0).
2-2 5’-phosphorylated ssDNA guides are dissolved to 100 ng/μl in 0.5x TE
buffer (PH 8.0). For each well of a 24-well plate, 200-250 ng NLS-NgAgo
plasmid and 100-300 ng guidesare diluted in 50 μl Opti-MEM (Gibco); 1.25 μ
l lipofectamine 2000 is diluted in 50 μl Opti-MEM.... 阅读全帖 |
|
C****n
发帖数: 79 | 31 谢谢分享您的经验。
我一直也是这么做的。Clontech 有设计网页，用起来也很方便。
overlap区域的话是20bp，也是自动设计的。理论上说长一些的话，效率会更高一些。
这个问题是刚换了实验室出现的。一个不同点是gel purification kit 。之前用的是
Qiagen的，后来嫌yield太低，换成Zymo的了。还有就是Nanodrop 不一样，但是应该不
会问题太大。
打算用Infusion Plus 自带的gel purification kit 试一下，然后也order了一个 NEB
的NEBuilder HiFi DNA Assembly。仍然不work的话，只能换其他方法了。
insertion
1 |
|
s*******e
发帖数: 740 | 32 Scientist/Associate Scientist of immunology
Responsibilities:
Primary immune cell isolation and culture
Antibody purification and characterization
DNA vector construction and cell transfection
Retrovirus production and cell transducing
Skills/Requirements
Experience in primary immune cell culture, immune cell-based assays, flow
cytometry, and cell sorting
Experience in retrovirus preparation and cell transducing
Experience in protein purification from yeast, E.coli and mammalian cells
Experience... 阅读全帖 |
|
m***c
发帖数: 637 | 33 招聘：世界500强上海研发中心诚聘以下物理化学生物医药海外留学人士。
If you are interested, please email your CV to d****[email protected] with the
position that you are applying as the title.
Assistant Research Scientist-Food Science
Assistant Research Scientist-Polymer
Assistant Research Scientist-Polymer
Assistant Research Scientist-Rheology in Deposition
Assistant Research Scientist-Water Purification
Assistant Research Scientist –Deposition
Product Development Technologist-Skin GDC护肤类产品开发研究(For Skin
Category)
Research Associate (Me... 阅读全帖 |
|
a***n
发帖数: 1500 | 34 獒蟆。。又见獒蟆。。
M
Institute
revolutio
and
Gradu
frameworks
purification,
In
the
c
Univer
frameworks
purification, |
|
s****2
发帖数: 4569 | 35 Director of DNA synthesis
Position Summary
We are seeking for a highly motivated scientist as a director of DNA
synthesis unit, leading a DNA synthetic group to establish DNA synthesis
facility and develop high quality DNA synthesis service at Wuxi Apptec. The
successful candidate will be expected to have extensive experience with DNA/
RNA synthesize facilities, oligo purification technology and a variety of
DNA synthesis products.
Required Qualifications
PhD in Biology, Chemistry, B... 阅读全帖 |
|
m***c
发帖数: 637 | 36 再转几个中国上海联合利华内部推荐招聘的位置给大家，尽量找联系人内部推荐。
中国上海联合利华内部推荐，招聘。
If you are interested, please email your CV to d****[email protected] with the
position that you are applying as the title.
Assistant Research Scientist-Polymer
Assistant Research Scientist-Rheology in Deposition
Assistant Research Scientist-Water Purification
Assistant Research Scientist –Deposition
Assistant Research Scientist-Polymer
Work in the Discover in R&D function for the research in Polymer Science
area and perform as an active project/team... 阅读全帖 |
|
s*******s
发帖数: 623 | 37 Job ID: GF2012A001
Posting Title: Research Scientist/Research Associate/Electrobiochemist
Brief Introduction of Gate Fuels Inc.:
“Imagination, Innovation, Implementation --- Enabled Technologies for a
Green Clean World”
Gate Fuels Inc. is a research start-up firm based in Blacksburg, VA, with
the goal of developing microbial and cell-free platform technologies that
enable low-cost production of biochemicals, biofuels and electricity. The
first platform is based on industrially-safe Bacillus subt... 阅读全帖 |
|
发帖数: 1 | 38 您好，不知道这个职位是否找到合适的人选，我的背景还是比较符合的，PhD期间做了
大量活性炭的工作，之间还在膜分离公司工作过四年有丰富的工程经验，请看我的简历
，如果您觉得还可以的话请与我联系，非常感谢！
Yuhe Cao
North Campus Drive 1400,
Agricultural and Biosystems Engineering,
South Dakota State University,
Brookings, SD 57007
(605) 592-0789
[email protected]/* */
Objective
Challenging research position in membrane science in a pharmaceutical, gas
adsorption, gas purification and separation, water treatment or biotech
company or university
Qualifications
• How fiber membrane fabr... 阅读全帖 |
|
发帖数: 1 | 39 您好，不知道这个职位是否找到合适的人选，我的背景还是比较符合的，PhD期间做了
活性物分离提取的工作，之间还在膜分离公司工作过四年有丰富的工程经验，请看我的
简历，如果您觉得还可以的话请与我联系，非常感谢！
Yuhe Cao
North Campus Drive 1400,
Agricultural and Biosystems Engineering,
South Dakota State University,
Brookings, SD 57007
(605) 592-0789
[email protected]/* */
Objective
Challenging research position in membrane science in a pharmaceutical, gas
adsorption, gas purification and separation, water treatment or biotech
company or university
Qualifications
• How fiber membrane fa... 阅读全帖 |
|
发帖数: 1 | 40 您好，不知道这个职位是否找到合适的人选，我的背景还是比较符合的，PhD期间做了
活性物分离提取的工作，之间还在膜分离公司工作过四年有丰富的工程经验，请看我的
简历，如果您觉得还可以的话请与我联系，非常感谢！
Yuhe Cao
North Campus Drive 1400,
Agricultural and Biosystems Engineering,
South Dakota State University,
Brookings, SD 57007
(605) 592-0789
[email protected]/* */
Objective
Challenging research position in membrane science in a pharmaceutical, gas
adsorption, gas purification and separation, water treatment or biotech
company or university
Qualifications
• How fiber membrane fa... 阅读全帖 |
|
c******m
发帖数: 884 | 41 替朋友发帖．
-------------------------------------------------------------------------
Sino-American Biotech: Multiple Managerial Bioprocess Position Openings in
China for a US-China joint venture company.
The client company is an existing company well established and joint-venture
with a US Biotech company about over 10 years ago. They were very
successful in supplying Chinese market for diagnostic and reagents in the
past years. Recently they decide to expend the business into biological
product R&D... 阅读全帖 |
|
z**m
发帖数: 48 | 42 请站内联系， 本人申请了两个职位，有关protein science, protein purification,
protein purification process development, 准备再申请合适的相关职位， 求内部
推荐
如果有幸得中， 定当酬谢 ( 给内部refer的兄弟姐妹，或者能做决定的manager,定当
重谢， 请站内联系）
多谢各位提携 |
|
d******n
发帖数: 29 | 43 1.
Towards large scale methods for the selective release of periplasmic human
cystatin C from E. coli
Biotechnology Techniques, Volume 9, Number 3 (1995), 179-184
2.
Optimization of an osmotic shock procedure for isolation of a protein
product expressed in E. coli.
Biotechnol Prog. 2003 Sep-Oct;19(5):1541-6.
3.
Optimization of BLyS production and purification from Escherichia coli.
Protein Expr Purif 39:237-246.
thanks a lot!
4.
High-level accumulation of a recombinant antibody fragment in the p... 阅读全帖 |
|
m***8
发帖数: 7 | 44 我是化学工程Ph.D， 已经毕业了, 研究方向是bioseparation process development,
找工作找了好几个月没有结果，非常气馁。 请问版上有在药厂做process development
方面的前辈吗？ 恳请帮忙内推一下，不甚感激。
其实已经申请过很多purification process engineer 的工作，但是投简历都石沉大海
。 现在Bayer Healthcare 又有一个Process Engineer (Purification)的职位跟我的
方向十分符合，请问在bayer的版友能不能帮我递个简历呢？ 非常感谢，有缘的话请您
吃饭。:)
我的联系方式： c************[email protected] |
|
l****o
发帖数: 32 | 45 Please apply for it online ASAP if you are interested in this opportunity
and I will recieve your resume right after that. Thanks.
Auto req 30057BR
Job Posting Title Grad Intern - Operations
Career Category College Job
Employee Subgroup Seasonal FT
Country (State/Region) United States - California
Location (City) US - CA - Thousand Oaks
Amgen Job Description Amgen’s 10 – 12 week internship program offers
meaningful project experiences that impact patient’s lives. This... 阅读全帖 |
|
S*****a
发帖数: 63 | 46 We are looking for an energetic and dynamic hands-on leader with initiative
and a
collaborative spirit to serve as a Group Leader in our protein chemistry lab
. This is a fantastic
opportunity to drive antigen/antibody expression, purification, and QA to
support
various therapeutic biologics programs while providing leadership to our
discovery efforts.
Responsibilities:
• Set up and run, in a hand-on way, a Protein Chemistry group to
express, purify, and characterize proteins, fusion pr... 阅读全帖 |
|
S****u
发帖数: 42 | 47 来自主题: Pharmaceutical版 - 无标题 Please apply from the Pfizer Website:
https://pfizer.wd1.myworkdayjobs.com/PfizerCareers/job/United-States---
California---Rinat/Scientist--non-PhD---Protein-Engineering_4692599
Pfizer -CID
Pfizer Cancer Immunology Discovery (CID) is a South San Francisco
biotechnology unit within the Oncology division of Pfizer Worldwide Research
& Development, dedicated to doing its part to fulfill Pfizer's mission of
working together for a healthier world. CID is dedicated to developing new
protein and cell-b... 阅读全帖 |
|
p*******2
发帖数: 56 | 48 俺前天进了[email protected]，准备捂到6月15号，赌结果。它有两个pending，一个随时给结果
，另一个要到12号或者15号。只要有一个get approved，股价就会再翻个翻，有两个
approved当然会更高了。如果都failed，股价就会又回到1毛。风险就这些，想玩的可
以适当关注，选择时机进入。
详情见下面：
1.) Nephros (OTC:NEPH): Price - $0.14, Market Cap - $5M, Revenue (ttm) - $1.
7M, Net Loss - ($5.4M)
The Nephros hemodiafiltration (HDF) system is designed to improve the
quality of life for End-Stage Renal Disease (ESRD) patients while addressing
the critical financial and clinical needs of the care provider. The Nephros
HDF system remove... 阅读全帖 |
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dy
发帖数: 12 | 49 February->februum(purgation,purification)
April->aperio(to open, to unveil)
May->Maia(daughter of Atlas & Pleione, mother of Mercury by Jupiter)
In Latin, the number 7 is septem, 8 is octo, 9 is novem, 10 is decem. July
was
at first Quinctilis(quinque=5), August was at first Sextilis(sex=6). |
|
dy
发帖数: 12 | 50 February->februum(purgation,purification)
April->aperio(to open, to unveil)
May->Maia(daughter of Atlas & Pleione, mother of Mercury by Jupiter)
In Latin, the number 7 is septem, 8 is octo, 9 is novem, 10 is decem. July
was
at first Quinctilis(quinque=5), August was at first Sextilis(sex=6). |
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