YeastMedia
YEPD (non-selection)-1% yeast extract-2% peptone-1.5% agar (if needed for plates)After autoclaving add glucose to 2% by adding 100 ml of 20% solution to each litre media.SYNTHETIC MEDIA (selective media)-0.67% yeast nitrogen base without amino acids (Difco) 0919 15 3-1.5% agar (if needed for plates). Also add 1 pellet of NaOH per litre of agar to avoid mushy plates.After autoclaving add 100 ml of 20% glucose (or......閱讀全文
Yeast－Media
YEPD (non-selection)-1% yeast extract-2% peptone-1.5% agar (if needed for plates)After autoclaving add glucose to 2% by adding 100 ml of 20% solution
Yeast－Media,－Solutions－and－Stocks
Yeast Media:Note: Synthetic complete medium can be prepared by adding media supplements (see below).Medium using 6.7 g yeast nitrogen base without ami
Raft－culture－media－(aka－Green’s－media)
Raft culture media (aka Green’s media)3 parts DMEM (including glutamine or glutamax)1 part Ham’s F125% FCSvarious supplements (not everybody uses the
Tissue－Culture－Media
We use two different kinds of media. Most cells are grown in DMEM. A few lymphoid cell lines are grown in RPMI. Cells grown in DMEM must be grown in a
Antibiotic－Concentration－in－Media
Antibiotics should be added to lower than 60 C broth, or filter sterilized: See note. "C'' refers to chromosomal resistance: "P" plasmid based
酵母培養基和相關試劑
Yeast Hartwell's Complete (HC) Media?(Gottschling Lab)??Yeast Complete Media???Yeast Media?(PMCI)??Yeast Media, Solutions and Stocks?(Donis Keller
Cell－Culture－Media－and－Solutions
Antimycotic/antibiotic media:To 1 liter of sterile RPMI 1640 with 2mM L-glutamine, add:165.0 ml fetal bovine serum, heat inactivated12.0 ml 200mM (100
Cell－Culture－Media－and－Solutions
Cell Culture Media and SolutionsDec. 18, 1990R. VeileAntimycotic/antibiotic media:To 1 liter of sterile RPMI 1640 with 2mM L-glutamine, add:165.0 ml f
Bacterial－Media－Solutions－and－Stocks
3 agar?(200 ml)Add 6 grams agar to 200 ml deionized water. Autoclave to sterilize.1.6 agar?(200 ml)Add 3.2 grams agar to 200 ml deionized water.Autocl
MEDIA－FOR－EMBRYO－CULTURE－AND－MANIPULATION
M16 Medium (Protocol obtained from Karen Austen-Reed from SS Tan Laboratory, Anatomy Department)For oocyte maturation and routine culture of mouse emb
Yeast－Lysates－for－Westerns
Cells are grown for 2-3 days as 1.5ml prep. under selection for the plasmid of interest. Spin cells down 2.6K for 5min.Resuspend in 1ml 0.25m NaOH/1%
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
Dropout－plates－for－yeast
Materials(Solutions are all available from the media room)200ml bottle of 2x SD200ml bottle of 4% agar -- make sure to sign it out40% glucoseCSM minus
yeast:Assaying－mating
SetupYou have yeast strains that are deficient in mating (eg Ste12 knockouts) and would like to test whether transforming them with a plasmid that con
Fast－Yeast－Transformation
Protocol: Fast yeast transformationAdd 50 μl carrier DNA to a 1.5 ml tube.scrap cells from plate and add to the carrier DNA.Add in the following order
Yeast－DNA－Prep
Protocolgrow up yeast culture to appropriate density (near saturation)spin 1.5 mls of culture for 1 min in microfuge and aspirate off supernatantresus
Yeast－Nuclei－Isolation
This method gives yeast nuclei which look nearly purified microscopically. Nuclei isolated in this way do not give active transcription extracts when
Modified－Yeast－Transformation
Inoculate cells from an overnight culture into 50 ml YEPD and incubate at 30°C with shaking. Typically, add 0.1 to 0.2 ml saturated culture in the eve
Method:－Removal－of－Yeast－Contamination－from－Lymphoblast－Cultures
Purpose:This method is advantageous for saving the occasional cultures that become contaminated. Yeast contaminated cultures will appear cloudy when s
TISSUE－CULTURE－STOCK－SOLUTIONS－AND－MEDIA
MS MEDIUM FOR ARABIDOPSISTo?990?ml?H2O?add: Sucrose?...........?10.0??g MOPS?..............??0.5??g Agar?..............??8.0??g Adjust?pH?to?5.7
Retos－Buffers－－Media－Book
08-15 BufferCompoundMWFinal concentration200 mlGlycine75.07?0.1 M1.5 g1 M MgCl2?10 mM2 mlKOHto pH 9H2Oto 200 mlAAcetic Acid 1 MCompoundMWFinal concent
Embryonic－limb－bud－culture－in－media
Early in embryonic development, the region of the chick embryo which is determined to form a limb first differentiates from the rest of the embryo1. T
Live－Cell－Imaging－of－Yeast
Live Cell Imaging of YeastDaniel R. Rines, Dominik Thomann, Jonas F. Dorn, Paul Goodwin and Peter K. SorgerINTRODUCTIONThe development of cloning vect
Decontamination－of－cells－from－the－yeast
I???? Destroy yeast1.???? Aspirate medium and wash cell in PBS.2.???? Incubate cells at 37oC for 5 min in non-diluted antibiotic-antimycotic.3.???? In
Plasmid－isolation－from－yeast
Pick colonies into 0.5ml of SD-Leu (or other appropriate SD medium)Vortex for 1minLeave to grow O/N for 18-24h at 30°C, 230-250rpm (best in 5ml bijou)
Yeast－Genomic－DNA－Prep
Grow 10ml YPD cultures o/n. Figure out cell density; inoculate 30 ml YPD and grow o/n so that cell density is approximately 2 x 108cells/ml the next m
Endy:Yeast－Colony－PCR
MethodUsing sterile pipette tips, transfer a 1 mm colony into 50 uL of 60 U/ml Zymolyase3 uL of 1 U/mL Zymolyase stock solution47 uL of waterIncubate
Blackburn:Yeast－Colony－PCR
OverviewThis is a quick and easy yeast colony PCR protocol that does not require zymolyase step.Updated Protocol:?Blackburn Lab: Quick and Easy Yeast
酵母培養
Streaking Yeast Stocks?(Donis Keller Lab)Very nice protoocol for yeast workLong-Term Storage of Yeast Stocks?(Donis Keller Lab)Yeast storage and reviv
Yeast－Cell－Cycle－by－Flow－Cytometry
ReagentsCold absolute ethanol.0.5 M Na citrate stock (filtered), 50mM diluted stock.10 mg/ml RNase A (Boil 10 mins, cool, filter and store at -20°C).4