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30% Acrylamide 100g Acrylamide 2.6g Bis H2O to 330 ml
40% Acrylamide 380g Acrylamide 20g BIS H2O to 1 liter filter sterilize
Acrylamide/BIS (Low) 0.98g BIS 200g Acrylamide H20 to 500ml
Acrylamide/BIS (not too High) 3.8g BIS 60g Acrylamide H20 to 200ml
Acrylamide/BIS (High) 2.0g BIS 30g Acrylamide H20 to 100ml
500X Ampicillin 25mg/ml in H20 Filter Sterilize and Aliquot
Bis Tris HL5 2.1g Bis Tris 10g Proteose Peptone 5g Yeast Extract 10g Glucose H20 to 1 Liter
CP 10ml 1M Hepes pH7.5 10ml 0.1M EDTA 79.5ml Glycerol H20 to 1 Liter
Cyclase Reaction Mix 1500ul 2M Tris-HCl pH 8.0 60mg PC 2.75mg ATP 3.0ml H20 freeze in 2ml aliquots
Cyclase Stop Solution 1g ATP 85mg cAMP 4g SDS 2M Tris-HCl to pH 7.5 H20 to 200ml
DB 5.36g Na2HPO4 · 7H20 2.72g KH2PO4 4ml 2M MgSO4 0.8ml 1M CaCl2 H20 to 4 Liters DB Agar – 1.5g/100ml
Denaturing Buffer 438.3g NaCl 100g NaOH H20 to 5 Liters
50X Denhardts 5g Ficoll 5g Polyvinyl Pyrrilidone 5g BSA Pentax Fraction V BSA H20 to 500ml Filter Sterilize and Aliquot
DEPC H20 Add 0.1% DEPC to H20 Shake well and Autoclave
Destain 4L H20 4L Methanol 800ml Glacial Acetic Acid
10X Dialysis Buffer 58.44g NaCl 12.1g Tris 3.72g EDTA pH to 8.0 H20 to 1 Liter
DM Plates with G418 10g Peptone 2g Dextrose 0.717g Na2HPO4 · 7H20 (0.38g Anhydrous) 1.45 KH2PO4 15g Agar H20 to 1 Liter Autoclave and Cool to 50° C Add 4ml G418 stock solution
10X DNA Gel Loading Buffer 20ml 50X TAE 40g Sucrose H20 to 100ml
DNAse I 1mg/ml in 0.15M NaCl / 50% glycerol
0.1M DTT 0.077g DTT/5ml H20 Aliquot 0.5ml in 1.5ml tubes
5X EB 3.7g CHAPS 5ml 1M Hepes pH7.5 77mg DTT 0.5ml 0.1M EDTA 40ml Glycerol H20 to 100ml
10X E Buffer A – 5.52g NaH2PO4/200ml H20 B - 10.72g Na2HPO4-7 H20/200ml H20 Add 85ml A + 15ml B pH to 6.1 with B H20 to 200ml Freeze in 25ml Aliquots
1X E Buffer 20 ml 10X E Buffer 50mM Sucrose 3.42g H20 to 200ml Freeze in Aliquots
0.5M EDTA 73g EDTA powder (NOT sodium salt) pH to 7.0 to get into solution
ERB 20ml 1M Tris-HCl pH7.5 10ml 0.5M EDTA 80mg DTT 1g Benzamidine 10mg each TPCK, TLCK, TAME 600ul PIC II 2ml PIC I Dissolve the following in 10ml EtOH then add slowly to the rest: 1g Phenanthroline 250mg CBZ – Phenylalanine 100mg PMSF
5X Formaldehyde Gel Buffer for RNA 0.2M MOPS pH7.0 50mM Sodium Acetate 5mM EDTA
Formamide (deionized) Add 5g Ag501-X8 20-50 mesh to 50ml Formamide Stir over night covered in fume hood Gravity filter
FM Medium Make 5 Liter according to package directions Add 0.085g NaHCO3 0.15g Streptomycin Autoclave Store in Cold room
G418 20mg/ml in 10mM Hepes pH7.2 Filter Sterilize and Aliquot
HL-5 80g Maltose or 40g Dextrose 40g Proteose Peptone 20g Yeast Extract 3.86g Na2HPO4 · 7H20 (2.04g Anydrous) 1.94g KH2PO4 0.12g Streptomycin H20 to 4 Liters
2M Imidizole 272g Imidizole pH to 7.5 H20 to 1 Liter
6% Instagel 420g Ultra Pure Urea 145ml 40% Acrylamide 100ml 10X TBE 405ml H20 Filter Sterilize Store in foil covered bottle (dark)
20X KK2 Buffer 46.2g KH2PO4 20g K2HPO4 H20 to 1 Liter
LB Broth
LB Plates – add 15g Agar/1L Top Agar – add 0.35g Agar/50ml
10X Ligation Buffer 500ul 1M Tris-HCl pH7.5 5.1mg ATP Grade I 100ul 1M MgCl2 50ul 10mg/ml BSA 0.2M DTT 350ul H20
Lower Gel Stock 108.9g Tris base 2.4g SDS pH to 8.8 H20 to 600ml
LP Medium 1g Lactose 1g Peptone 0.27g KH2PO4 0.35g Na2HPO4 · 2H20 OR 0.279g Na2HPO4 Anydrous H20 to 1 Liter
Lysis Buffer with PICs 5.0mg PMSF, 1g Phenanthroline, and 250mg CBZ-Phen dissolved in 5ml EtOH 1g Benzamidine 10ml PIC I 1ml PIC II 100mg TPCK 20mg TLCK 100mg TAME
Maleate Buffer 11.6g Maleic Acid 8.76g NaCl 7g NaOH pH to 7.5 H20 to 1 Liter Add 0.2ml DEPC
2M Mg2+ Stock 24.6g MgSO4 · 7 H20 20.4g MgCl2 · 6 H20 H20 to 100 ml Autoclave
Neutralization Buffer 605g TRIS 438.3g NaCl pH to 8.0 H20 to 5 Liters
NZCYM 10g NZ Amine 5g NaCl 1g Casamino Acids 5g Yeast Extract 2g MgSO4 H20 to 1 Liter
Oligo Prehybe 30.3ml 20X SSC 60ml H20 1.34g Na2HPO4 · 2H20 pH to 6.8 with HCl add 10ml 50X Denhardts 0.1mg/ml boiled Salmon Sperm DNA for probing add 100mg/ml Dextran Sulfate
Oligo Prewash 1.25ml 2M Tris-HCl pH8.0 29g NaCl 1ml 0.5M EDTA 2.5ml 20% SDS H20 to 500ml
Oligo Wash Buffer 3M Tetramethylammonium chloride 50mM Tris-HCl pH8.0 2mM EDTA 1mg/ml SDS
20X PBS 320g NaCl 8g KCl 46g Na2HPO4 · 7H20 8g KH2PO4 0.8g Na azide H20 to 4 Liters
10X PCR Buffer 2.5ml 2M Tris-HCl pH8.0 1.87g KCl 750ul 1M MgCl2 2.5ml 2% gelatin filter sterilize
50% PEG 250g PolyEthyleneglycol (MW=8000) H20 to 500ml Stir for 1-2 hours
10X Phosphate Buffer 27.2g KH2PO4 53.6g Na2HPO4 · 7H20 H20 to 4 Liters
PIC I – 1000X 25mg Antipain 50mg Leupeptin 25mg Aprotin H20 to 25ml
PIC II – 1000X 1mg/ml chymostatin 1mg/ml pepstatin dissolve in DMSO
10X Ponceau S 2g Ponceau S 30g Trichloracetic acid 30g Sulfosalyclic acid H20 to 100ml
3M Potassium Acetate pH 5.0 147.2g Potassium Acetate 57.5ml Glacial Acetic Acid H20 to 500ml
Proteinase K 20mg/ml in H2O
QBT 21.95g NaCl 5.2325g MOPS 75ml EtOH 0.75ml Triton X100 pH to 7.0 H20 to 500ml
QC 29.22g NaCl 5.232g MOPS 75ml EtOH pH to 7.0 H20 to 500mL
QC 73.05g NaCl 25ml 2M Tris-HCl pH8.0 150ml EtOH pH to 8.5 H20 to 500ml
Random Prime Stop 100ul 20% SDS 100ul 0.5M EDTA 40ul 10mg/ml tRNA 760ul sterile H20
10X Low Salt Restriction Buffer 200ul 1M Tris-Hcl ph7.5 100ul 1M MgCl2 100ul 10mg/ml BSA 600ul H20
10X Medium Salt Restriction Buffer 200ul 1M Tris-Hcl ph7.5 100ul 1M MgCl2 100ul 5M NaCl 100ul 10mg/ml BSA 500ul H20
10X High Salt Restriction Buffer 200ul 1M Tris-Hcl ph7.5 100ul 1M MgCl2 300ul 5M NaCl 100ul 10mg/ml BSA 300ul H20
RLB 54.8g Sucrose 5ml 1M Tris-HCl pH7.5 0.508g MgCl2 · 6 H20 5ml Triton X-100
RNAse A 10mg/ml in: RNAse A Buffer Heat to 100° C for 20 minutes Let cool slowly to room temperature
RNAse A Buffer 1ml 1M Tris-HCl pH7.5 0.3ml 5M NaCl H20 to 100ml Autoclave
RP Prehybe 5ml 20% SDS 20ml 5M NaCl 50ml Formamide 10g Dextran Sulfate H20 to 100ml
10X Running Buffer 121.3g TRIS base 576g Glycine 40g SDS H20 to 4 Liters
2.5X Sample Buffer 25ml Glycerol 12.5g DTT 7.5g SDS 15.6ml 1M Tris-HCl pH7.5 5mg Bromophenol Blue H20 to 100ml
SM Stock Solution 5.8 NaCl 2g MgSO4 · 7 H20 (0.97g Anhydrous) 50ml 1M Tris-HCl pH7.5 5ml 2% gelatin H20 to 1 Liter
SM Plates 4.62g KH2PO4 2.0g K2HPO4 20g Dextrose 20g BactoPeptone 2g Yeast Extract H20 to 2 Liters 20g Agar/1L
SM/3 Plates 4.62g KH2PO4 2.0g K2HPO4 6.6g Dextrose 6.6g BactoPeptone 0.66g Yeast Extract H20 to 2 Liters 20g Agar/1L
SM/5 Plates 4.62g KH2PO4 2.0g K2HPO4 4g Dextrose 4g BactoPeptone 0.4g Yeast Extract H20 to 2 Liters 20g Agar/1L
3M Sodium Acetate 123g Sodium Acetate pH to 5.2 with Glacial Acetic Acid H20 to 500ml
20X SSC 701.2g NaCl 352.8g Sodium Citrate H20 to 4 Liters
Stain 4L H20 4L Methanol 800ml Glacial Acetic Acid 9g Brilliant Blue
STET 40g Sucrose 2.5ml Triton X+100 2.5ml 2M Tris-Hcl pH8.0 50ml 0.5M EDTA H20 to 500ml
Super 2X YT
Superbroth 24g Yeast Extract 12g Tryptone 5ml Glycerol H20 to 900ml
Sequence Stop Solution 45% Formamide 20mM EDTA 0.05% Bromophenol blue 0.05% Xylene Cycnol FF
SOB Medium 20g Bacto Tryptone 5g Yeast Extract 0.5g NaCl 1.86g KCl pH to 7.0 H20 to 1 Liter
Sonicated Salmon Sperm 10mg/ml 0.3g DNA 30ml H20 Stir for a long time in ice 0.5ml aliquots at -20° C
50X TAE 242g TRIS base 57.1ml Glacial Acetic Acid 100ml 0.5M EDTA H20 to 1 Liter
10X TBE 121.1g Tris base 55g Boric Acid 7.4g Na2EDTA H20 to 1 Liter Filter Sterilize
20X TBST 193.6g TRIS base 640g NaCl pH to 7.6 80ml Tween-20 H20 to 4 Liters
TE 5ml 1M Tris-HCl pH7.5 1ml 0.5M EDTA H20 to 500ml
TEG 12.5ml 1M Tris-Hcl pH 7.5 10ml 0.5M EDTA 4.5g Glucose H20 to 500ml
TES 6.25ml 2M Tris pH8.0 10ml 0.5M EDTA 75g Sucrose H20 to 500ml
10X Transfer Buffer 120g TRIS base 576g Glycine H20 to 4 Liters
1X Transfer Buffer dilute 10X stock 1/10 and then add 250ml/L of Methanol
1M Tris-HCl 141g TRIS base in 800ml H20 pH to 7.5 H20 to 1 Liter
2M Tris-HCl 242g TRIS base in 700ml H20 pH to 8.0 H20 to 1 Liter
10X Tris Phosphate 108g Tris base 40ml 0.5M EDTA 15.1ml 85% Phosphoric Acid
Upper Gel Stock 36.3g Tris base 2.4g SDS pH to 6.8 H20 to 600ml
Uracil 0.4g Uracil H20 to 200ml at 56° C Sterile filter
5X Wash Buffer 121g TRIS base 175g NaCl 4g NaN3 10ml NP-40 66.6ml BSA pH to 8.0 H20 to 4 Liters
YPD medium 10g Yeast Extract 20g Peptone 20g Glucose H20 to 1 Liter Plates – 20g agar/1L
2X YT 16g Bacto Peptone 10g Yeast Extract 5g NaCl H20 to 1 Liter
This page was last edited 06/26/2003 |