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Isolating Postimplantation Embryos: Early Somite-Stage Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4366

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause postimplantation defects. This protocol describes a method for isolating early somite-stage embryos (~8.5 days post coitum [dpc]). - [Read Isolating Postimplantation Embryos: Early Somite-Stage Protocol]

Isolating Postimplantation Embryos: Late Primitive-Streak-Stage Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4364

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause postimplantation defects. This protocol describes a method for isolating late primitive-streak-stage embryos (~7.5 days post coitum [dpc]). - [Read Isolating Postimplantation Embryos: Late Primitive-Streak-Stage Protocol]

Isolating Postimplantation Embryos: Prestreak-Stage Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4362

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations that cause post-implantation defects. This protocol describes a method for isolating prestreak-stage embryos (~5.5 days post coitum [dpc]). - [Read Isolating Postimplantation Embryos: Prestreak-Stage Protocol]

Isolation and Culture of Mouse Bone Marrow-Derived Macrophages Protocol - http://www.scienceboard.net/resources/protocols.asp?action=article&protocol_id=572&criteria=

Category: Immunology: Mononuclear Cell Isolation Protocols

Mononuclear phagocyte progenitor cells derived from femoral and tibial bone marrow are propagated in the presence of M-CSF. This macrophage growth factor is secreted by L929 cells and is used in the form of L929 cell conditioned medium. The progenitor cells proliferate and differentiate through monoblast, promonocyte and monocyte stages before maturing to macrophages. At this time the cells become firmly adherent to the culture vessel. - [Read Isolation and Culture of Mouse Bone Marrow-Derived Macrophages Protocol]

Isolation and Freezing of Primary Mouse Embryonic Fibroblasts (MEF) For Feeder Plates - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4482

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the isolation of fibroblasts from mouse embryos. Mouse embryonic fibroblast (MEF) cells are used as a feeder layer for the culture of mouse embryonic stem (ES) cells to help maintain them as pluripotent stem cells. The inhibition of ES-cell differentiation provided by the MEF feeders appears to be due to their production of leukemia inhibitory factor (LIF). - [Read Isolation and Freezing of Primary Mouse Embryonic Fibroblasts (MEF) For Feeder Plates]

Isolation and Growth of Primary Cell Cultures from Mouse Protocol - http://www.stanford.edu/dept/EHS/prod/researchlab/bio/docs/Palmer_Bio_protocol.pdf

Category: Mouse Protocols: Mouse Cell Culture Protocols

The procedures involve the isolation and growth of primary cell cultures from rodent and human tissue as well as the use of viral vectors for the introduction and expression of mammalian genes in cells in culture and in live rodents. - [Read Isolation and Growth of Primary Cell Cultures from Mouse Protocol]

Isolation and handling of primary mouse embryonic fibroblasts (MEFs) - http://www.molgen.mpg.de/~rodent/MEF_protocol.pdf

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: MEF Protocols Mouse Embryonic Fibroblasts

Embryonic stem cells are usually grown on a layer of mitotically inactivated primary mouse embryonic fibroblasts to promote growth and prevent differentiation. Boris Greber - [Read Isolation and handling of primary mouse embryonic fibroblasts (MEFs)]

Isolation and Purification of Tetrahymena Nuclei Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4500

Category: Tetrahymena Protozoa Protocols

Protocol describes how to allow the isolation of nuclei from all stages of the Tetrahymena life cycle in high yield with a high degree of purity. This method gives highly purified populations of both micronuclei and macronuclei. - [Read Isolation and Purification of Tetrahymena Nuclei Protocol]

Isolation of Adult Mouse Cardiac Myocytes from One Heart Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000125.pdf?pid=PP00000125

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

This procedure describes the isolation and culture of adult mouse cardiac myocytes from one heart. The isolation routinely yields approximately 1 million rodshaped myocytes per heart. - [Read Isolation of Adult Mouse Cardiac Myocytes from One Heart Protocol]

Isolation of Adult Mouse Cardiac Myocytes from One Heart Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000125.pdf?pid=PP00000125

Category: Mouse Protocols: Mouse Cell Culture Protocols

Protocol describes the isolation and culture of adult mouse cardiac myocytes from one heart. The isolation routinely yields approximately 1 million rodshaped myocytes per heart. - [Read Isolation of Adult Mouse Cardiac Myocytes from One Heart Protocol]

Isolation of Adult Mouse Cardiac Myocytes from Two or More Hearts Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000126.pdf?pid=PP00000126

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

This procedure describes the isolation and culture of adult mouse cardiac myocytes from two or more hearts. Includes modifications for the digestion of two or more hearts in the same procedure and subsequent pooling of myocytes derived from the multiple hearts. The isolation procedure is performed by one or more technicians and routinely yields approximately 1 million rod-shaped myocytes per heart. - [Read Isolation of Adult Mouse Cardiac Myocytes from Two or More Hearts Protocol]

Isolation of Arabidopsis cDNAs by Complementation in Yeast - http://www.arabidopsis.org/cshl-course/12-yeast.html

Category: Plant Biology Protocols: Arabidopsis Nucleic Acid Protocols

Includes: Isolation of Arabidopsis LEU2 cDNAs by complementation of the yeast leu2 mutation; Recovery of plasmid DNA from yeast cells; Analysis of Arabidopsis cDNAs that complement a yeast leu2 mutation; Preparation of library DNA from the Lacroute cDNA library; Preparation of Yeast Media. - [Read Isolation of Arabidopsis cDNAs by Complementation in Yeast]

Isolation of Arabidopsis Nuclei and Measurement of Gene Transcription Rates Protocol - http://www.natureprotocols.com/2007/01/25/isolation_of_arabidopsis_nucle.php

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol for the isolation of Arabidopsis nuclei and measurement of gene transcription rates using nuclear run-on assays. Plant materials are ground in hexylene glycol-based buffers and highly enriched nuclear fractions are obtained using Percoll density gradients. Standard and small-scale protocols are presented, along with a tested method for nuclear run-on assays. The entire process may be completed within 3 days. - [Read Isolation of Arabidopsis Nuclei and Measurement of Gene Transcription Rates Protocol]

Isolation of BAC DNA from Large-scale Cultures Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4007

Category: BACs Bacterial Artificial Chromosome Protocols

The procedure for isolation of BAC DNA is scaled-up to accommodate 500-ml cultures, which, on average, yield 20-25 µg of purified BAC DNA. - [Read Isolation of BAC DNA from Large-scale Cultures Protocol]

Isolation of BAC DNA from Small-scale Cultures Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4006

Category: BACs Bacterial Artificial Chromosome Protocols

BAC DNAs are prepared from 5-ml cultures of BAC-transformed cells by a modification of the standard alkaline lysis method (Preparation of Plasmid DNA by Alkaline Lysis with SDS: Minipreparation). The yield typically varies between 0.1 and 0.4 µg of BAC DNA. - [Read Isolation of BAC DNA from Small-scale Cultures Protocol]

Isolation of Chloroplasts from Spinach Leaves Protocol - http://homepages.gac.edu/~cellab/chpts/chpt8/ex8-1.html

Category: Plant Biology Protocols: Photosynthesis and Respiration Protocols

Protocol describing the isolation of chloroplasts from spinach leaves. - [Read Isolation of Chloroplasts from Spinach Leaves Protocol]

Isolation of Dendritic Cells Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E66369ADF1DF1962F01D3DA073183AB&objectid=6674A1FCBE1A61EE40A5E3EE8FB368F5

Category: Immunology: Mononuclear Cell Isolation Protocols

Presents two methods for preparing dendritic cells (DCs), a highly specialized type of antigen-presenting cell (APC). The first method involves the isolation of DCs from mouse spleen, resulting in a cell population that is highly enriched in accessory cell and APC function. A support protocol for collagenase digestion of splenocyte suspensions is described to increase the yield of dendritic cells. The second method involves generating large numbers of DCs from mouse bone marrow progenitor cells. - [Read Isolation of Dendritic Cells Protocol]

Isolation of DNA Fragments from Polyacrylamide Gels by the Crush and Soak Method Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot2936

Category: Nucleic Acid Electrophoresis Protocols: DNA Electrophoresis Protocols: DNA Electrophoresis in Polyacrylamide Gels Protocols

The "crush and soak" method, which works best for DNAs <1 kb in size, can be used to recover both singleand double-stranded DNAs from polyacrylamide gels. The yield of eluted DNA varies from <30% to >90% depending on the size of the DNA fragment. - [Read Isolation of DNA Fragments from Polyacrylamide Gels by the Crush and Soak Method Protocol]

Isolation of DNA from Mammalian Cells by Spooling Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4037

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: DNA Isolation from Mammalian Cells Protocols

Procedure is used to prepare DNA simultaneously from many different types of samples or tissues. Although the DNA is generally too small (approx. 80 kb) for efficient construction of genomic DNA libraries, it gives excellent results in Southern hybridizations and PCRs. Cultured aneuploid mammalian cells (2 x 107, e.g., HeLa cells) yield 100 µg of DNA in a volume of 1 ml. - [Read Isolation of DNA from Mammalian Cells by Spooling Protocol]

Isolation of DNA from Mammalian Cells Grown in 96-well Microtiter Plates Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3695

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: Genomic DNA Isolation Protocols

Protocol for isolation of DNA from mammalian cells grown in 96-well microtiter plates. Each well yields sufficient genomic DNA for several standard PCRs or for analysis in a single lane of a Southern hybridization. - [Read Isolation of DNA from Mammalian Cells Grown in 96-well Microtiter Plates Protocol]

Isolation of DNA from Mouse Tail Biopsies - http://www.med.umich.edu/tamc/tDNA.html

Category: Transgenic Mouse Protocols: Mouse Tail DNA Extraction Genotyping

Isolation of DNA from Mouse Tail Biopsies. University of Michigan Transgenic Animal Model Core, Transgenic Animal Web. - [Read Isolation of DNA from Mouse Tail Biopsies]

Isolation of DNA from Strawberries and Raspberries - http://www.science-projects.com/onionDNA.htm

Category: DNA Protocols: DNA Extraction Protocols

Also works for onions. Duration ~40 minutes. DNA Isolation with very easily accessible household solutions. Great for school classes. science-projects.com - [Read Isolation of DNA from Strawberries and Raspberries]

Isolation of Fibroblasts from Chicken Embryos Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4475

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

This protocol describes the isolation of fibroblasts from chicken embryos. - [Read Isolation of Fibroblasts from Chicken Embryos Protocol]

Isolation of Fibroblasts from Mammalian Embryos Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4474

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

This protocol describes the isolation of fibroblasts from mouse, rat, and hamster embryos. - [Read Isolation of Fibroblasts from Mammalian Embryos Protocol]

Isolation of gametes of Sea Urchins - http://www.swarthmore.edu/NatSci/sgilber1/DB_lab/Urchin/Urchin_gamete.html

Category: Developmental Biology: Sea Urchin Developmental Biology

Protocol for the isolation of gametes of Sea Urchins. Cebra-Thomas. Swathmore. - [Read Isolation of gametes of Sea Urchins]

Articles (1-5 of 5)

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

Acid Guanidinium Thiocyanate RNA Isolation Phenol Chloroform Extraction

A single step RNA isolation protocol using Phenol Chloroform Extraction and Acid Guanidinium Thiocyanate. This RNA isolation method uses the fact that guanidinium thiocyanate can simultaneously lyse the cells and inactive cellular RNAses during the initial RNA isolation step allow a single step in the method.

DNA Fragment Purification from Polyacrylamide Gels Protocol

A simple protocol for the purification of DNA fragments from Polyacrylamide Gels.

DNA Ligation Protocol

The DNA Ligation protocol described here contains the steps required to join together using ligase enzyme both plasmid DNA and insert DNA fragments in order to create a new plasmid. This new ligated plasmid can be transformed after into competent bacteria to produce DNA for mini, midi or maxi-prep isolation.

Electrotransformation of BMH 81-17mut S for Isolating Site-Directed dsDNA Mutants

This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.

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