CountingEScellChromosomes
(original reference: "tissue culture made easy" by Christian LaMantia from the Magnuson lab)1) Plate cells onto gelatinized plates (35 or 60 mm) without feeders and culture 24 hours. A 1:4 split is best for most lines.2) Culture for four hours in the presence of colcemid (3.125 µl of 10 µg/ml colcemid (Gibco # 15212-012) per ml of medium). 2ml per 35 mm and 4ml per 60 mm.3) Remove media and place it in a 15......阅读全文
Counting－ES－cell－Chromosomes
(original reference: "tissue culture made easy" by Christian LaMantia from the Magnuson lab)1) Plate cells onto gelatinized plates (35 or 60 mm) witho
Cell－counting/plating
OverviewCount the number of viable cells in a culture via dilution plating. The following assumes E. coli cells but it should apply to any type of cel
Cell－counting－with－an－hemacytometer.
Accuracy of manual counts with an hemacytometer depend on:accurate mixing of the sample (no bubbles!)number of chambers countednumber of cells counted
Cell－Counting/－LiveDead－Discrimination
This is a microscopy based application for definitive discrimination of live and dead cells. Trypan Blue exclusion notoriously over estimates the numb
Human－Embryonic－Stem－(ES)－Cell－Protocols——General－notes－on－ES－cell－culture
hES media has a two week shelf life.hES cells should be cultured in 4, 6, 24, 48, 96 well plates. Growing cells in flasks is not recommended because i
ES－Cell－Culture－and－Manipulation
MediaHigh glucose DMEM (-pyruvate, -glutamine)20% Heat-inactivated Fetal calf serum (can vary by cell type, be sure!!)1X l-glutamine1X Penicillin/stre
ES－and－TS－cell－freezing/thawing
实验概要ES and TS cell freezing/thawing.主要试剂ES cell freezing medium (2x) 2x ES cell freezing medium should be made up fresh each time it is to be
ES－and－TS－cell－freezing/thawing
Needed:ES cell freezing medium (2x)2x ES cell freezing medium should be made up fresh each time it is to be used, and should comprise freshly prepared
Human－Embryonic－Stem－(ES)－Cell－Protocols——Thawing－Human－ES－cells
Remove Human ES cells from liquid nitrogen storage tank. Fill out a freeze/thaw form.Thaw cryovial by gently swirling in waterbath until only a small
Human－Embryonic－Stem－(ES)－Cell－Protocols——Freezing－Human－ES－Cells
Collagenase cells for approximately 7 minutes at 37 °C (until edges of colonies are curling up).With a 5 ml pipet, gently pipet and scrape colonies f
Human－Embryonic－Stem－(ES)－Cell－Protocols——Splitting－Human－ES－cells－on－MEFs
based on splitting onto one plateWarm collagenase IV split media to 37 °C in a water bath.Aspirate media off of cell culture plate.Add the following a
Human－Embryonic－Stem－(ES)－Cell－Protocols—Splitting－Human－ES－cells－onMatrige
based on splitting onto on plateWarm collagenase media to 37°C in a water bath.Aspirate media off of cell culture plate.Add the following amount of co
ES－Cell－Culture－and－Manipulation3
Care and Handling of Feeder Layer CellsSTO/SNL cells are a derivative of the standard STO cell line. They are transformed with a LIF producing plasmid
ES－Cell－Culture－and－Manipulation2
Picking ES cell clonesOne or two days before picking colonies prepare 24-well plates of feeders. You can also use alternate protocols that utilize 96-
MEDIA－AND－SOLUTIONS－REQUIRED－FOR－ROUTINE－ES－CELL－CULTURE
Media UsedTo prepare 100 ml mediumDMEM80 mlFCS15 mlNon-essential amino acids (100x)1 mlPen/strep (5,000 1U/ml, 5000 ug/ml)1 mlL-Glutamine 200 mM1 mlNu
细胞计数试剂盒(Cell－Counting－Kit)操作方法
一、制作标准曲线（测定细胞具体数量时）1、先用细胞计数板计数所制备的细胞悬液中的细胞数量，然后接种细胞。2、按比例（例如：1/2比例）依次用培养基等比稀释成一个细胞浓度梯度，一般要做3-5个细胞浓度梯度，每组3-6个复孔。3、接种后培养2-4小时使细胞贴壁，然后加CCK试剂培养一定时间后测定OD值，
Human－Embryonic－Stem－(ES)－Cell－Protocols——Media－and－Reagents
Serum Free Media for human ES cells on MEFs: can last for 7-10 daysFinal ConcentrationAmount for 250ml Stock solution80% DMEM-F12200ml20% KO Serum Rep
Human－Embryonic－Stem－(ES)－Cell－Protocols——Embryonic－Bodies
Let human ES cells grow until the colonies are large and the cells are pretty piled up - about the time when you would normally split or even a day pa
Using－a－Counting－Chamber
Using a Counting ChamberFor microbiology, cell culture, and many applications that require use of suspensions of cells it is necessary to determine ce
Human－Embryonic－Stem－(ES)－Cell－Protocols——Matrigel－Aliquoting－and－Plating
Aliquoting Matrigel:Day one:Put the sterilized tip box (either 200 ml or 1000 ml tips), sterilized eppendorf tube container, and appropriate pipettor
FIXATION－OF－PLANT－METAPHASE－CHROMOSOMES
Reagents (a) Metaphase arresting agents: Choose one of the following (Note 2) and shake vigorously to aerate before putting in living plant materi
In－Situ－Hybridization－to－Somatic－Chromosomes－in－Drosophila
In Situ Hybridization to Somatic Chromosomes in DrosophilaAbby F. DernburgINTRODUCTIONIn situ hybridization was originally developed as a technique fo
Horizontal－Transfer－of－Supernumerary－Chromosomes－in－Fungi
Several species of filamentous fungi contain so-called dispensable or supernumerary chromosomes. These chromosomes are dispensable for the fungus
Mapping－Protein－Distributions－on－Polytene－Chromosomes－by－Immunostaining2
12. Hold the salivary glands at the common duct with tweezers.Transfer the glands to a drop of fixing solution on a siliconizedcoverslip. 13. Incubate
杨芃原：Exploration－of－human－chromosomes－by－biomass－spectrometry
复旦大学化学系 杨芃原教授 2014年4月26日，首届全国质谱分析学术研讨会在北京西郊宾馆盛大开幕。来自复旦大学的杨芃原教授带来了题为《Exploration of human chromosomes by biomass spectrometry based
Mapping－Protein－Distributions－on－Polytene－Chromosomes－by－Immunostaining1
INTRODUCTIONThe formidable size and structure of polytene chromosomes allowmapping of chromosomal protein distributions at very high resolution.This p
Electroporation－of－ES－cells
Cells are routinely passaged two days prior to electroporating. Usually one 10 cm plate at approximately 80% confluency will provide enough cells for
KARYOTYPING－ES－CELLS
An actively growing culture of cells is required, i e 2 - 3 d ES cell culture. The total number of cells needs to be between 106 - 107 cells.N B Read
ES细胞分化培养实验——培养皿ES细胞集落
实验材料ES 细胞单一胚胎样体仪器、耗材细菌培养皿ES 分化培养基实验步骤1. 准备下列试剂和材料：悬滴培养的 ES 细胞单一胚胎样体100 mm 细菌培养皿ES 分化培养基2. 在 100 mm 细菌培养皿中加 10 ml 预热的分化培养基。3. 从温箱中取出培养 2 天的悬滴培养物。小心翻转培养
DIRECT－AND－SHORTTERM－PROCEDURE－FOR－HARVESTING－BONE－MARROW－CHROMOSOMES
I. PurposeTo identify chromosome anomalies in hematopoietic cells. Used especially for chromosome studies for hematological disorders such as preleuke