Isolationofmouseembryos
1. Sacrifice impregnated mouse.2. Dissect out the uterus of the mouse. Pulling up on the uterus with one set of forceps,use another to tear the mesometrium away from the uterus.3. Remove the embryos from the uterus. Separate them into one or two embryo fragments.4. Using fine forceps, separate the muscular wall of the uterus, Reichert's membrane and visceral yolk sacfrom the actual embryo.......閱讀全文
Isolation－of－mouse－embryos
1. Sacrifice impregnated mouse.2. Dissect out the uterus of the mouse. Pulling up on the uterus with one set of forceps,use another to tear the mesome
Isolation－of－Zebrafish－embryos
Zebrafish will mate and deposit fertilized eggs on the bottom of the tank at 'dawn'. They can be accustomed to lay at any convenient time by k
Isolation－and－growth－of－mouse－primary－myoblasts
Isolation of limb muscle from neonatal mice1.?Neonatal mice by decapitation or CO2 inhalation.2.?Rinse the limbs with 70% ethanol and remove them wi
ISOLATION－OF－PRIMARY－MOUSE－EMBRYO－FIBROBLASTS
You will need:13.5 day pregnant mouse (we use MTK NEO inbred white mice)2 sets sterile instrumentsone containing a pair of curved forceps and a pair o
Mouse－B－cell－isolation－with－magnetic－beads
實驗概要Isolate B cells from total mouse cells實驗步驟?Praparation:1.?????? Enough MACS; put RPMI into water bath.2.?????? Put some MACS in 15 ml tube or 1.5
胚胎干細胞培養
Media and Solution required for ES Cell Culture?(Bowtell Lab)???Routine Culturing of ES Cells?(Bowtell Lab)??Routine Splitting and freezing of cells?(
轉基因
DNA PreparationGene TransferEmbryo TransferTransgenic IdentificatioinOthersTransgenic Outline?(University of Michigan Transgenic Animal Model Core)Thi
Fixation－of－Embryos
MEMFA Fix10xMEMFA Salts1 part 10x MEMFA salts1 M MOPS1 part 37% formaldehyde20mM EGTA8 parts water10mM MgSO410x salts can be autoclaved and stored. Tu
RNA提取
RNA提取（主要內容如下）Tips for Handing RNA?Total RNA IsolationmRNA IsolationrRNA IsolationOthersQ & A posted in the Method Forum??Basic Procedures for Handing
Mouse－Spleenectomy
OUTLINEPROTOCOLGeneral anesthesia1. inject i.p. 250μl/mouse of the?ANESTHETIC mixture?< wait for 2-10min for the anesthetic effect>2. for verify the a
Wholemount－staining－of－embryos
Fix embryos in formalin or MEMFA for one hour at room temperature with mixing. Rinse with TBS, replace with methanol, store at -20oC.Rehydrate by slow
Preparation－of－Mouse－Neutrophils
實驗步驟Mice：8-16 weeks old malePrewarm buffer to room temperatureBuffer A : Ca2 ?/Mg2 ‐free Hank’s buffered saline solution [HBSS; Invitrogen, Grand Isla
Complete－Mouse－Necropsy
EuthanasiaEuthanasia and mouse necropsies require prior IACUC approval. The mode of euthanasia should be chosen which minimizes pain or distress to th
Mouse－keratinocyte－cultures
PRIMARY MOUSE KERATINOCYTE CULTURESIsolation of epidermal keratinocytes from neonatal mice is based on the protocol of Dlugosz et al.,?Methods Enzymol
Preparation－of－Mouse－Neutrophils
實驗概要Preparation of Mouse Neutrophils實驗步驟Mice：8-16 weeks old malePrewarm buffer to room temperatureBuffer A : Ca2 ?/Mg2 ‐free Hank’s buffered saline so
Pathology－and－Autopsy－of－a－mouse
if mouse is found deadif mouse is a newborn or < 1 wk of ageIf mouse is alive or moribund and above one week of ageRoutine dissectionFormalin fixation
Sectioning－stained－embryos.
The protocols for plastic and wax sections as used by the Vize lab. These protocols work, but they have not been optimized. If anyone has better proto
In－vitroculture－of－early－chick－embryos
1. Use sterile technique. Prewarm Howard's Ringers in petri dish andagar/albumin culture dish to 37oC.2. Crack 2-day egg into large sterile petri
TUNEL－PROCEDURE－IN－BOVINE－EMBRYOS
Materials8% (w/v) paraformaldehyde stock solution:??Dissolve 8 g of powdered paraformaldehyde in 100 ml water. Heat and stir (55-60 C – do not go high
TUNEL－PROCEDURE－IN－BOVINE－EMBRYOS
Materials8% (w/v) paraformaldehyde stock solution:??Dissolve 8 g of powdered paraformaldehyde in 100 ml water. Heat and stir (55-60 C – do not go high
Obtaining－Embryos:－hCG－Injection
Ovulation is induced in X. tropicalis by injection of human chorionic gonadotropin (hCG). X. tropicalis requires a much smaller dose than does X. laev
多功能胚胎激光破膜儀（一）
Laser-assisted derivation of human embryonic stem cell lines from IVF embryos after preimplantation genetic diagnosis.T. Turetsky, E. Aizenman, Y. G
Immunofluorescent－staining－of－Sea－Urchin－embryos
1. Transfer fixed embryos to microfuge tubes. Allow to settle for 10 minutes.Gently remove most of the liquid.2. Add 100 ul antibody to one tube and 1
真核細胞總RNA的制備（Total－RNA－Isolation）2
Total RNA Isolation?Guanidine-based isolation Objective: ?To obtain total RNA from zebrafish embryos. Required Materials ?Denaturing Solution
RNAse－A－Treatment－of－Mouse－Cells
IntroductionRNAse A treatment of permeabilized cells followed by immunostaining is a method which allows to show if the localization of a protein into
Culturing－Mouse－Embryonic－Fibroblasts
MaterialsTrypsin (Gibco 25200-023)3T3 Medium:? 500 mL DME (Invitrogen) + 50 mL FBS (Hyclone) + 5 mL 100x Pen/Strep2x Freezing Medium: 3T3 Medium + 20%
Mouse－Compete－Blood－Counts
Materials:?250 μL of fresh mouse blood in plastic tubes containing EDTA.?RBC lysis buffer?(388 mM NH4Cl, 29.7 mM NaHCO3, 25 μM Na2EDTA)20.75g. NH4CL2.
Chick－or－Mouse－embr...
實驗概要The following procedure describes the procedure for whole mount staining of chick or mouse embryo’s. A similar procedure could be used for sta
小鼠feeder細胞分離
You will need:13.5 day pregnant mouse (we use MTK NEO inbred white mice)2 sets sterile instrumentsone containing a pair of curved forceps and a pair o
Effect－of－Cyclopamine－on－Xenopus－laevis－embryos
Cyclopamine and jervine, teratogens derived from the skunk cabbage?Veratrum californicum, are known to induce holoprosencephaly in chick embryos when