GrowingOvernightCultures
1. Place 2 mL of the appropriate sterile medium in a 13 mm yellow-capped culture tube. If more culture is needed, place up to 5 mL in a 16 mm green-capped culture tube. Large amount of culture should be made in growth flasks, using up to 25 flask volume in medium (e.g., no more than 30 mL in a 125 mL flask, or 250 mL in a 1000 mL flask).2. Touch a single colony with the end of an applicator stick and hold it in your ......阅读全文
Growing－Overnight－Cultures
1. Place 2 mL of the appropriate sterile medium in a 13 mm yellow-capped culture tube. If more culture is needed, place up to 5 mL in a 16 mm green-ca
Transformation－of－E.－coli－by－Electroporation
实验概要 Electrocompetent bacteria are prepared by growing cultures to mid-log phase, washing the bacteria extensively at low temperature, and
Growing－cells
No two cell lines behave exactly the same, so you must learn the peculiarities, or personality, of each of the cell lines with which you work. Irrespe
Preparing－Overnight－Bacterial－Culture
Materials:Sterile LB medium (Luria-Bertani Medium) with or without antibiotic:water 500 mlbacto-tryptone 10 gbacto-yeast extracts 5 gsodium chloride 1
Plastic－dishes－for－growing－cells
There are two kinds of dishes used to grow tissue culture cells.Those that are designed for adherent cells have been treated chemically to promote cel
Lambda－DNA－Preparation
Lambda DNA PreparationThis is a plate method that gives very good yield for cloning. I have combined the Promega and Maniatis protocols.Solutions T-TY
Method:－Maintaining－Lymphoblastoid－Cell－Lines
Method: Maintaining Lymphoblastoid Cell LinesJune 10, 1990Rosalie VeilePurpose:To grow lymphoblastoid cells for permanent storage and for DNA extracti
ORGANOTYPIC－KIDNEY－CULTURES
-embryos are dissected from timed-pregnant mice from 11.5 d.p.c. to 13.5 d.p.c.-metanephroi and associated ureteric buds are microdissected and placed
Hippocampal－Neuron－Cultures
实验概要The protocol provides a method of hippocampal neuron cultures.主要试剂Begin by timing the pregnant mouse at E17-E19 days of gestation. Have ready the
Mouse－keratinocyte－cultures
PRIMARY MOUSE KERATINOCYTE CULTURESIsolation of epidermal keratinocytes from neonatal mice is based on the protocol of Dlugosz et al., Methods Enzymol
ORGANOTYPIC－LIMB－CULTURES
-embryos are dissected from timed-pregnant mice from 10.5 - 11.5 d.p.c.-limb buds are microdissected and placed in holding medium (L15 medium suppleme
Preserving－yeast－cultures
Short term storageYeast cultures are stable for 1-2 weeks when refrigerated. Petri dishes should be sealed or in plastic bags.Medium term storageYeast
Cryopreservation－of－cell－cultures
1. Examine all flasks by inverted-phasemicroscopy. Cultures used for preservation should be grown free of antibiotics, show no signs of microbial cont
A－novel－method－of－growing－fungi－for－DNA－extraction
Preparation of fungi for DNA extraction typically involves growing cultures in liquid culture in Erlenmeyer flasks, Roux bottles or even microfuge tub
Tissue－Culture－Methods1
I. TYPES OF CELLS GROWN IN CULTURETissue culture is often a generic term that refers to both organ culture and cell culture and the terms are often us
Dissociated－Cultures－of－Cerebellar－Neurons
Dissociated Cultures of Cerebellar NeuronsHank Dudek (617-355-4735)Protocolisolate cerebella (ÒCbÓ)cut off head into plate with HHGNhold nose with lar
Transfer－of－Eukaryote－Suspension－Cultures
MaterialsFibroblast suspension cultureTissue culture laminar flow hoodMedia appropriate to culture line usedDisposable pipettes (10 ml and 1.0 ml)Disp
Primary－Cultures－fo...
实验概要The following protocol provides a method of primary cultures for IHC – viability assays.实验步骤1. Preparation of primary mesencephalic cultures 1)
AMNIOTIC－FLUID－CULTURES－ON－COVERSLIPS
I. Purpose:Amniotic fluid may be used for prenatal diagnosis of aneuploidy or other structural abnormalities. II. Culture Procedure:A. Aseptic techniq
AMNIOTIC－FLUID－CULTURES－ON－COVERSLIPS
实验概要 AMNIOTIC FLUID CULTURES ON COVERSLIPS主要试剂Solutions:Colcemid working solution: 10 mcg/ml Colcemid in Hank's Balanced Salt Solution, sto
酵母染色体沉淀分析方法
ABSTRACTThis protocol describes a method for the detection of proteins bound to specific regions of chromatin in yeast. There are many variations of t
Growing－feederindependent－embryonic－stem－cells§
We use feeder-independent ES cell lines derived from the 129/Ola strain of mice (Nichols et al., Development 110, p.1341, 1990). These cells are easy
Fungal－Genomic－DNA－Extraction
OverviewHigh throughput of many fungal isolates can be achieved by growing axenic cultures in either (a) 1.5mL microfuge tubes, half full with liquid
Fungal－Genomic－DNA－Extraction
OverviewHigh throughput of many fungal isolates can be achieved by growing axenic cultures in either (a) 1.5mL microfuge tubes, half full with liquid
SDS－Whole－Cell－Extracts
-Use sterile technique and sterile solutions in steps 1 to 3.-1. Using a saturated starter culture, inoculate 25 to 30 ml of appropriate media in a 12
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
Competent－agro－prep－for－electroporation
day 11. Start 75 mL overnight cultures of agro (strain GV3101 C58C1 Rifr pMP90 Gmr, Koncz & Schell) in YEP in 250 mL baffle flasks.2. Grow at 28 °
TISSUE－FIBROBLAST－CULTURES－FOR－CHROMOSOME－ANALYSIS
I. Purpose:A: Skin tissue may be used for chromosome analysis in special cases when the results from peripheral blood are inconclusive, e.g. suspected
Rodent－Retinal－Ganglion－Cell－Cultures
实验概要Central neurons lose the ability for axonal regrowth during development and typically do not regenerate their axons following axotomy once the
SOLID－TUMOR－CULTURES－FOR－CHROMOSOME－ANALYSIS
I. Purpose:A. Samples of solid tumors or lymph nodes may be sent from patients with cancer. These samples should be processed directly and also set up