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![ProZyme/Glyko®2-AA低聚甘露糖6（Man6）[GKSA-104]/GKSA-104/100 pmol](/images/prozyme/201712/GKSA-104_2AA_Man6_2f6d0246-7bd7-49ea-b7af-cb5b42b0c30b_large.png?v=1442230102)
常用试剂和培养基
蚂蚁淘
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Sterilewaterbottles
================ 蚂蚁淘在线 ================ 免责声明：本文仅代表作者个人观点，与本网无关。其创作性以及文中陈述文字和内容未经本站证实，对本文以及其中全部或者部分内容、文字的真实性、完整性、及时性本站不做任何保证或承诺，请读者仅作参考，并请自行核实相关内容 版权声明：未经蚂蚁淘在线授权不得转载、摘编或利用其他方式使用上述作品。已经经本网授权使用作品的，应该授权范围内使用，并注明“来源：蚂蚁淘在线”。违反上述声明者，本网将追究其相关法律责任。200ml,500ml,or950mlwater.Autoclave.
0.05MCaCl2(per200ml)CaCl2·2H2O1.5gAdd198mlwater.Autoclave.
0.5MCaCl2(per200ml)CaCl2·2H2O15.0gAdd188mlwater.Autoclave.
0.1MMgSO4(per200ml)MgSO4(anhydrous)2.4gAdd200mlwater.Autoclave.
10%MSG(for200ml)Glutamicacid(Na)20.0gAdd188mlwater.Autoclave.
40%Glucose(per200ml)Glucose(dextrose)80.0gAddwaterto200ml(~145-150ml).Autoclave.
10%CAA(per200ml)CaseinAcidHydrolysate20.0gAdd186mlwater.Autoclave.
10%Yeast(per200ml)Yeastextract20.0gAdd150-160mlwater.Autoclave.Add150ulß-mercaptoethanolafterautoclaving.
20XTryptone/NaCl(per200ml)Tryptone40.0gNaCl20.0gAdd190mlwater.Autoclave.Add150ulß-mercaptoethanolafterautoclaving.
10XM9stock(for200ml)Na2HPO4(anhydr.)12.0gKH2PO46.0gNaCl1.0gNH4Cl2.0gAdd196mlH2O.Autoclave.Note:leaveoutNH4Cltoprepare10XM9-N.
10XM9-Nstock(for200ml)Na2HPO4(anhydr.)12.0gKH2PO46.0gNaCl1.0gAdd196mlH2O.Autoclave.Note:leaveoutNH4Cl!
100XP(for200ml)KH2PO418.1gK2HPO45.0gpHto6.3with10NKOH!Add192mlH20.Autoclave.
1XSM(for200ml)NaCl1.16gMgSO40.20gGelatin(2%)1.00mlor20mgAdd190mlH2O,thenadd:Tris(1M,pH7.5)10.0mlAutoclave.
2%Gelatin(per500ml)Gelatin10gAdd500mlwater.Autoclave.
20%Succinate(for500ml)Succinicacid100gKOH90gAddto400mlicecoldwater.MeasurepHto6.4,filtersterilize.
10XTBE(electrophoresisbuffer)Trisbase108.0gNa2EDTA9.3gBoricAcid55.0gBringvolumeto1Lwithwater.Usedoubledistilledwater.pHshouldbe8.3;ifnotadjustdropwisewithconc.NaOH.
Wateragarstock(for190ml)Agar3.0gAdd190mlH2O.Autoclave,storein60Cwaterbath.
Ragarstock200ml500mlTryptone........2.0g5.0gNaCl............1.0g2.5gAgar............3.0g7.5gAddH2O.........194.0ml485.0mlAutoclave,storein60Cwaterbath.
LBagar200ml1000mlTryptone......2g10gNaCl..........1g5gYeastextract.1g5gAgar..........3g15gAddH2O......193ml990mlAutoclave,storein60Cwaterbath.LBagarcanalsobemadebyadding10ml10%yeastextractand10ml20XTryptone/NaClto190mlWA.
NZ200ml500mlNZamine....2.0g5.0gNaCl........1.0g2.5gMgSO4(anhydrous)..0.2g0.5gAdd10MNaOH(1drop/100ml)afteraddingwater.Autoclave.Foragarmedia:Agar........2.4g6.0gStirbeforeandafterautoclaving.
GYPCtoWAstock:200ml1000mlGlucose(40%).....2.0ml10.0mlYeastextract(10%)4.0ml20.0ml100XP............2.0ml10.0mlCAA(10%).........4.0ml20.0mlSuccinate(20%)....1.0ml5.0mlAddappropriateantibiotics.Pourplates.
ORSminimal200ml1000ml10XM9.........10ml50mlSuccinate(20%).2ml10mlMSG(10%)......2ml10mlGlucose(40%)..1ml5mlMgSO4(0.1M)..1ml5mlCaCl2(0.5M)..200ul1ml1000Xvitamins.200ul1mlWater..........184ml920mlOmitsuccinateandadd10mlglucoseforstoragemedium.
E.coliminimal200ml1000ml10XM9..........20ml100mlMgSO4(0.1M)...2ml10mlCaCl2(0.5M)...0.2ml1ml1000Xvitamins..0.2ml1mlGlucose(40%)...2ml10ml
XCminimal200ml1000ml10XM9..........5.0ml25mlMgSO4(0.1M)...1.0ml5mlCaCl2(0.5M)...0.2ml1ml200X7aastock..0.2ml1mlGlucose(40%)...0.5ml2.5mlGlycerol(50%)..4.0ml20ml
Nifplates10XNifbuffer100ml10Xtraceelements100ml1000Xvitamins1mlSterileddH2O799mlAgarose6gSuccinate(20%)20ml(Succinateisalready0.4%fromthatin10Xnifbuffer)
10XNifbuffer1000mlK2HPO434.02gKH2PO457.06gSuccinate(20%)200.00mlpHshouldbe6.4after10Xdilution.
1000XORSvitaminsNicotinicacid2.0mg/mlPantothenate1.0mg/mlBiotin0.2mg/mlStir,filtersterilize.
TraceelementsMgSO4(0.1M)25mlCaCl2(0.5M)5mlNa2MoO4(1mg/ml)5mlFeCl3(1mg/ml)5mlWater460ml
YEM(per500ml)K2HPO40.5gMgSO47H2O0.2gNaCl0.1gMannitol10.0gYeastextract0.4gAgar15.0gpHto6.9
YEMsubstitute(per200ml)To180mlWAstock:10XM98.0mlMannitol(20%)20.0mlYeastextract(10%)1.6ml
SM10selectivemedia(forstorage)10XversatileMg100mlThiamine5mlThreonine5mlLeucine5mlGlucose(40%)10mlWater875ml
AminoacidsSM10stocks:Thiamine(0.337%)Threonine(0.71%)Leucine(0.79%)Klebsiella:Histidine(0.31%)
10XversatileMg(for200ml)MgSO42.0mlThiamine(1mg/ml)2.0mlCaCl20.2ml
200XPLT(for200ml)Proline9.20gLeucine1.60gThiamine0.67gAutoclave.
MgPLT10XversatileMg100ml200XPLT5ml40%glucose10mlWater885ml
Zbuffer(lacZassays)200ml1000mlNa2HPO47H2O....3.22g16.10gNaH2PO4H2O......1.10g5.50gKCl..............0.15g0.75gMgSO4(0.1M)....1.00ml0.20mlß-mercaptoethanol.2.70ml0.54mlFiltersterilize.
ONPGstock4mg/mlinphosphatebuffer(pH7)
Phosphatebuffer(pH7.0)per100ml:Na2HPO47H2O1.64gNaH2PO4H2O1.38g
0.2MNa2CO3(100ml)Na2CO3(anhydr.)10.6g
Jensen"sMedium(Sesbania)10XMacronutrients(1000ml):K2HPO43H2O2.6gNaCl2.0gMgSO40.97gCaCl210.0gFeCl36H2O1.4gKNO33.0g
Jensen"sMedium(Sesbania)1Xsolution(1000ml):K2HPO43H2O0.20gNaCl0.20gMgSO47H2O0.20gCaHPO41.00gFeCl36H2O0.10gorFeEDTA0.26gKNO3.(onlyfor10X)3.00g
Jensen"sMedium(1942)seeVincent1970p.75.perLiter:K2HPO43H2O0.2gNaCl0.2gMgSO47H2O0.2gCaHPO41.0gFeCl36H2O0.1gorFe-EDTA0.26gDilute1:5whenused.15mldilutedmedium/pouch.
1000XMicronutrients(200ml)H3BO3572mgMnCl2406mgZnSO4H2O44mgCuSO45H2O16mgNaMO4BH2O18mgCo2+20mgNi2+20mg
Nodulegrindingbuffer(500ml)50mMKH2PO4(pH7.6),0.15MNaClK2HPO4(1M)3.25mlKH2PO4(1M)21.75mlNaCl4.38gmAddto450mldistilledwater,pHto7.6,andadjustvolumeto500ml.
RapidynegermicideForworkingsolution:Diluteconcentratedstockat0.6ml/L.
1MTris-Cl(500ml)To400ml,add:Trisbase60.5gAddconc.HCltopHbringtovolume.
TrisbuffermixturesforpHpHTrisHClTrisbase7.0.923.0777.1.905.0957.2.90.107.3.865.1357.4.84.167.5.82.187.6.78.227.7.74.267.8.67.337.9.63.378.0.50.50
SouthernTransfersolutionsI.0.25NHCl:21.5mlconc.HCl(11.6N)to1litervolume.II.0.5MNaOH,1.5MNaCl:87.7gNaCl,20.0gNaOHto1litervolume.III.1.0MTris,1.5MNaCl:121.1gTrisbase,87.7gNaCl,pHto8.0withconc.HCl(~50ml),adjustto1litervolume.IV.20XSSC:175.3gNaCl,88.2gNa3citrate,pHto7.0withconc.HCl(~1drop),adjustto1litervolume.
Random-primeroligonucleotidelabellingMix:9.00ulDNA(notmorethan200ng)1.25ulhexamersBoil,thenplaceonice,thenadd:5.0ulHEPESbuffer5.0ulnucleotides[in250mMTris(pH8),25mMMgCl2,50mMß-mercaptoethanol]1.0ulBSA(40mg/ml)0.5ulKlenow5.0ul-32PCTPLetreactiongoatroomtemperatureforatleast4hours,orovernight.
MSMEDIUMFORARABIDOPSIS
To990mlH2Oadd:Sucrose...........10.0gMOPS..............0.5gAgar..............8.0gAdjustpHto5.7with1MKOH.Autoclave.Whencooledaddsterilestocksolutionsof:KNO3...............20.0ml940mM(95.00g/L)NH4NO3.............20.0ml1000mM(80.04g/L)KH2PO4.............12.5ml100mM(13.60g/L)MgSO47H2O.........15.0ml100mM(24.65g/L)CaCl27H2O.........10.0ml300mM(44.00g/L)Glycine............2.67ml10mMNicotinate.........0.4ml10mMThiamin............0.02ml10mMPyridoxineHCl.....0.24ml10mMMyoinositol........5.5ml100mMFeSO47H2O........1.0ml(2.780g/100ml)Na2EDTA...........1.0ml100mM(3.720g/100ml)MnSO44H2O.........1.0ml100mM(1.690g/100ml)ZnSO44H2O.........3.0ml10mM(0.288g/100ml)H3BO3.............1.0ml100mM(0.618g/100ml)KI.................0.5ml10mM(0.166g/100ml)CuSO45H2O........0.01ml10mM(0.170g/100ml)Na2MoO42H2O......0.1ml10mM(0.242g/100ml)CoCl26H2O........0.01ml10mM(0.238g/100ml)B5MEDIUMFORARABIDOPSIS
To990mlH2Oadd:Sucrose...........20.0gMOPS..............0.5gAgar..............8.0gAdjustpHto5.7with1MKOH.Autoclave.Whencooledaddsterilestocksolutionsof:KNO3...............26.3ml940mM(95.00g/L)NaH2PO4H2O........11.0ml100mMMgSO47H2O.........10.0ml100mM(24.65g/L)(NH4)2SO3..........10.0ml100mMCaCl27H2O.........3.3ml300mM(44.00g/L)Myoinositol........5.5ml100mMThiamine...........3.0ml10mMNicotinate.........0.8ml10mMPyridoxineHCl.....0.48ml10mM10XMicronutrients.10.0ml(seebelow)Sequestrene330Fe.10.0ml(0.28g/100ml)ForCallusInducingMedium(CIM)add:2,4-D.....................0.50mlKinetin...................0.05mlForShootInducingMedium(SIM)add:IAA.......................0.15ml6-dimethylaminopurine.....5.00ml
10XMicronutrients(per100ml)MnSO44H2O.........0.6ml100mM(1.690g/100ml)ZnSO44H2O.........10.07ml10mM(0.288g/100ml)H3BO3.............4.9ml100mM(0.618g/100ml)KI.................4.5ml10mM(0.166g/100ml)CuSO45H2O........2.3ml10mM(0.170g/100ml)Na2MoO42H2O......1.0ml10mM(0.242g/100ml)CoCl26H2O........1.0ml10mM(0.238g/100ml)
LB(Luria-Bertani)BrothBacto-Tryptone10gBacto-YeastExtract5gNaCl5gH2Oupto1LIfdesired,adjustthepH7.5withNaOH.
CA(CompleteAgar)CaseinHydrolysate10gYeastExtract5gK2HPO44gAgar15gH2Oupto1L
StabAgarNutrientBroth10gNaCl5gAgar6gH2Oupto1LMixwellandautoclavefor15minat121C.
S.O.C.(fromBRL)1X100ml2%Bacto-Tryptone2.0g0.5%YeastExtract0.5g10mMNaCl2.92g2.5mMKCl3.73g20mMMgCl220.2g20mMMgSO424.6g20mMGlucose9.0gMixabove,bringto100ml,andfilterthroughasterile0.45µmfilter.Storethestocksolutionat-20Cor-70C.
BacillusThuringiensisMediumComponentG/LGlucose3.0(NH4)2SO42.0YeastExtract2.0K2HPO43H2O0.5MgSO4*7H2O0.2CaCl2*2H2O0.08MnSO4*4H2O0.05Addcomponenetstodistilledwater.MixthroughlyandbringpHto7.3.Autoclavefor15minat15psi--121°C
