Contributor:Zappe,H Related:E.coligrowth Yeasttryptonebroth/agar,2xYT foragarplates agar--15g autoclave. Contributor:Zappe,H Related:manyprotocols Storage:4C Ammoniumacetate CanbeusedasSolutionIIIforplasmidpreparation. Contributor:Zappe,H Related:SDS-PAGEgels Storage:Makefreshweekly Ammoniumpersulphate(APS) Contributor:Zappe,H Related:manyprotocols Storage:-20C Ampiciliin(Sigma,sodiumsalt),1000x filtersterilizeandstorealiquotsat-20C dilute1/1000forfinalconcof100mg/ml Contributor:Zappe,H Related:manyprotocols Storage:-20C ATP(0.1M) AdjusttopH7.0with0.1MNaOH(spottingafewmlonpHpaper) Freeze0.1mlaliquots Contributor:Zappe,H Related:Bal31shortening Storage:4C Bal31reactionbuffer,5x Allthecomponents,excepttheglycerol,arecombinedina10mlsterilintube.Thetubeisthenfilledtothe10mlmarkwithsterileglycerol.Mixbygentlyshakingthetube.ThevolumemarksonthesterilintubeareusedasitisnotpossIBLetoaccuratelyPipette5mlofundilutedglycerol. Contributor:Zappe,H Related:GelbasedDNAmanipulation TAbuffer ?? Contributor:Zappe,H Related:Gelelectrophoresis Tris-boratebuffer(5x) DonotstoreTris-borateinaglassbottle. Contributor:Zappe,H Related:manyprotocols TEBuffer Contributor:Zappe,H Related:manyprotocols Storage:4C Calciumchloride(1M),CaCl2 autoclave Contributor:Zappe,H Related:manyprotocols Care: Chloroform:Isoamyl(24:1) Contributor:Zappe,H Related:- Cysteine Contributor:Zappe,H Related:DNAdigestion Storage:-20C Restrictionenzymedilutionbuffer Contributor:Zappe,H Related:DNAdigestion Storage:-20C Restrictionenzymedilutionbuffer(10x) SaltconcentrationmM Contributor:Zappe,H Related:DNAdigestion Storage:-20C SmaIrestrictionenzymereactionbuffer(10x) Allthecomponents,excepttheglycerol,arecombinedina10mlsterilintube.Thetubeisthenfilledtothe10mlmarkwithsterileglycerol.Mixbygentlyshakingthetube.Thevolumemarksonthesterilintubeareusedasitisnotpossibletoaccuratelypipette5mlofundilutedglycerol. Contributor:Zappe,H Related:BacterialDNAextraction Cellsuspensionbuffer: Ifthecellsaregoingtobephysicallydisrupted(Frenchpressorglassbeads),thendonotaddthesucrose. Contributor:Zappe,H Related:BacterialDNAextraction Lysingsolution(2X): AddproteinaseK(50*lofa20mg/mlstocksolutionper10ml2Xlysingsolution)andthe*-mercaptoethanoltotheamountoflysingsolutionneeded.TheotherComponentscanbepremixedasastocksolution. Contributor:Zappe,H Related:manyprotocols Storage:-20C Dithiothreitol(DTT) filtersterilizeandstorealiquotsat-20C Contributor:Zappe,H Related:manyprotocols EDTA(Ethylene??) EDTAwillnotdissolveuntil+/-pH8.0 autoclave Contributor:Zappe,H Related:manyprotocols Care: Ethidiumbromide(EtBr) Contributor:Zappe,H Related:manyprotocols Ethanol(EtOH)Recipeforyeasttryptonemedium
Keywords:Zappe;YT2x;Yeasttryptone;E.coli
YeasttryptonemediumComponentstockconcfinalconc./1lTryptone--16gYeastextract--10gNaCl--5g
Recipeforammoniumacetate
Keywords:ammoniumacetate;Zappe
AmmoniumacetateComponentstockconcfinalconc./200ml/1lAmmoniumacetate-7.5M289gwater--to200ml
RecipeforAmmoniumpersulphate
Keywords:Zappe;APS;ammoniumpersulphate
AmmoniumpersulphateComponentstockconcfinalconc./0.5mlAPS10%(w/v)0.05gwater0.5ml
Recipefoampicillin
Keywords:ampicillin;Zappe
AmpicillinComponentstockconcfinalconc./20ml/1lAmpicillin-100mg/ml2gwater--to20ml
RecipeforATP
Keywords:ATP;Zappe
ATPComponentstockconcfinalconc./0.8mlml/1lATP-0.1M60mgNaOH0.1M-adjusttopH7.0water--to0.8ml
RecipeforBal31nucleasereactionbuffer
Keywords:Zappe;Bal31;DNAshortening;buffer
Recipes:BAL-31reactionbuffer(5X).Componentstockconcfinalconc./10mlTris-HCl,pH8.01M0.1M1mlCaCl21M60mM0.6mlMgCl21M60mM0.6mlEDTA,pH8.00.5M5mM0.1mlNaCl5M3M6mlWater-1.7ml
RecipeforBal31nucleasedilution/storagebuffer
Keywords:Zappe;Bal31;DNAshortening;buffer
Recipes:BAL-31dilution/storagebufferContributor:Zappe,HRelated:Bal31shorteningStorage:-20CBal31dilutionandstoragebufferComponentstockconcfinalconc./10mlTris-HCl,pH8.01M20mM0.2mlCaCl21M5mM50mlMgCl21M5mM50mlEDTA,pH8.00.5M1mM20mlNaCl5M0.1M0.20mlWater-5.58mlglycerol44%(v/v)4.40mlRecipeforTAbuffer
Keywords:TAbuffer;ligation;Zappe;electrophoresis
Recipes:TAbufferComponentstockconcfinalconc./10mlTris-HCl,pH8.01MNaAcl5MWater???-5.58ml
RecipeforTris-boratebuffer
Keywords:Zappe;TB;electrophoresis;gel
TBbufferComponentstockconcfinalconc./1000mlTris-base54gBoricacid27.5gEDTA,pH8.00.5M20ml
RecipeforTEBuffer
Keywords:TEbuffer;Babb
TEbufferComponentstockconcfinalconc./10ml/100ml/1lTris,pH8.01M10mM0.1ml1ml10mlEDTA,pH8.00.5M1mM20ml0.2ml2ml??
Recipeforcalciumchloride
Keywords:Zappe;calciumchloride;cacl2
CalciumchlorideComponentstockconcfinalconc./100ml/1lCaCl2.2H20-1M14.7gwater--to100ml
Recipeforchloroform:isoamylalcoholmixture
Keywords:chloroform:isoamylalcoholmix;Babb
Chloroform:isoamylalcoholmixcautionComponentstockconcfinalconc./10ml/100mlChloroform---96mlIsoamylalcohol---4ml
Recipeforcysteine
Keywords:cysteine
Recipes:CysteineComponentstockconcfinalconc./10mlCysteine0.1M1mM0.1mlWater-to4.69ml
DNAdigestion:restrictionenzymedilutionbuffer
Keywords:Zappe;DNA;digestion;restriction
RestrictionenzymedilutionbufferComponentstockconcfinalconc./10mlTris-HCl,pH7.51M10mM0.1mlNaCl5M50mM0.1mlbmercapto-ethanol10mM7mlGelatin10mg/ml100mg/ml0.1mlwater5.3mlglycerol44%(v/v)4.4ml
DNAdigestion:restrictionenzymecorereactionbuffer
Keywords:Zappe;DNA;digestion;restriction
Restrictionenzymecorebuffers(10X).Componentstockconc050100150Tris-HCl,pH7.51M1ml1ml1ml1mlNaCl5M01ml2ml87.7mgBSA(albumin)10mg/ml1ml1ml1ml1mlMgCl21M1ml1ml1ml1mlDTT0.5M0.2ml0.2ml0.2ml0.2mlwater2.4ml1.4ml0.4ml2.4mlglycerol44%(v/v)4.4ml4.4ml4.4ml
DNAdigestion:SmaIrestrictionenzymereactionbuffer
Keywords:Zappe;DNA;digestion;restriction;SmaI
SmaIrestrictionenzymereactionbufferComponentstockconcfinalconc./10mlTris-HCl,pH8.01M0.1M1mlKCl5M0.2M2mlMgCl21M0.2M2mlDTT0.5M0.2ml0.2mlwater0.4mlglycerol44%(v/v)4.4ml
RecipeforDNAextractioncellsUSPensionbuffer
Keywords:Cellsuspensionbuffer;DNAextraction;Zappe
CellsuspensionbufferComponentstockconcfinalconc./100ml/1l/10mlTris-HCl(pH8.0)1M10mM1mlEDTA(pH8.0)0.5M1mM0.2mlsucrose-0.35M12gwater--to100ml
RecipeforDNAextractionlysingsolution
Keywords:Lysingsolution;DNAextraction;Zappe
LysingsolutionComponentstockconcfinalconc./100ml/1l/10mlTris-HCl(pH8.0)1M100mM10mlEDTA(pH8.0)0.5M20mM4mlNaCl5M0.3M6mlSDS25%(w/v)2%8mlb-mercaptoethanol-2%(v/v)2mlproteinaseK20mg/ml100mg/ml0.5mlwater--to100ml
RecipeforDTT
Keywords:Zappe;DTT;dithiothreitol
Dithiothreitol(DTT)Componentstockconcfinalconc./20ml/1lDTT-1M3.09gsodiumacetate,pH5.2--to20ml
RecipeforEDTA
Keywords:EDTA;ethyl???;Zappe
EDTA(ethyl??)Componentstockconcfinalconc./400ml/1lEDTA-0.5M93.05gNaOH--+/-10gwater--to400ml
Recipeforethidiumbromide
Keywords:Zappe;EtBr;ethidiumbromide
Ethidiumbromide(EtBr)neverbreathdust-mutagenComponentstockconcfinalconc./10ml/1lEtBr-10mg/ml0.1gwater--to10ml
Recipeforethanolstock
Keywords:ethanol;EtOH
Ethanol(EtOH)Componentstockconcfinalconc./500ml/1lEtOH-70%350mlwater--150ml
