Recipe for yeast tryptone medium
Keywords: Zappe; YT 2x; Yeast tryptone; E. coli
Yeast tryptone medium
Contributor: Zappe, H
Related: E. coli growth
Yeast tryptone broth/agar, 2xYT
Component stock conc final conc. /1l
Tryptone - - 16g
Yeast extract - - 10g
NaCl - - 5g
for agar plates
agar - - 15g
autoclave.
Recipe for ammonium acetate
Keywords: ammonium acetate; Zappe
Ammonium acetate
Contributor: Zappe, H
Related: many protocols
Storage: 4 C
Ammonium acetate
Component stock conc final conc. / 200 ml /1l
Ammonium acetate - 7.5M 289g
water - - to 200ml
Can be used as Solution III for plasmid preparation.
Recipe for Ammonium persulphate
Keywords: Zappe; APS; ammonium persulphate
Ammonium persulphate
Contributor: Zappe, H
Related: SDS-PAGE gels
Storage: Make fresh weekly
Ammonium persulphate (APS)
Component stock conc final conc. / 0.5ml
APS 10%(w/v) 0.05g
water 0.5ml
Recipe fo ampicillin
Keywords: ampicillin; Zappe
Ampicillin
Contributor: Zappe, H
Related: many protocols
Storage: -20 C
Ampiciliin (Sigma, sodium salt), 1000x
Component stock conc final conc. / 20 ml /1l
Ampicillin - 100 mg/ml 2g
water - - to 20ml
filter sterilize and store aliquots at -20 C
dilute 1/1000 for final conc of 100 mg/ml
Recipe for ATP
Keywords: ATP; Zappe
ATP
Contributor: Zappe, H
Related: many protocols
Storage: -20 C
ATP (0.1M)
Component stock conc final conc. / 0.8ml ml /1l
ATP - 0.1M 60mg
NaOH 0.1M - adjust to pH 7.0
water - - to 0.8ml
Adjust to pH 7.0 with 0.1M NaOH (spotting a few ml on pH paper)
Freeze 0.1ml aliquots
Recipe for Bal31 nuclease reaction buffer
Keywords: Zappe; Bal31; DNA shortening; buffer
Recipes: BAL-31 reaction buffer (5X).
Contributor: Zappe, H
Related: Bal31 shortening
Storage: 4 C
Bal31 reaction buffer, 5x
Component stock conc final conc. / 10ml
Tris-HCl, pH 8.0 1M 0.1M 1ml
CaCl2 1M 60mM 0.6ml
MgCl2 1M 60mM 0.6ml
EDTA, pH 8.0 0.5M 5mM 0.1ml
NaCl 5M 3M 6ml
Water - 1.7ml
Recipe for Bal31 nuclease dilution/storage buffer
Keywords: Zappe; Bal31; DNA shortening; buffer
Recipes: BAL-31 dilution/storage buffer Contributor: Zappe, H Related: Bal31 shortening Storage: -20 C Bal31 dilution and storage buffer Component stock conc final conc. / 10ml Tris-HCl, pH 8.0 1M 20mM 0.2ml CaCl2 1M 5mM 50 ml MgCl2 1M 5mM 50 ml EDTA, pH 8.0 0.5M 1mM 20 ml NaCl 5M 0.1M 0.20ml Water - 5.58ml glycerol 44%(v/v) 4.40ml
All the components, except the glycerol, are combined in a 10 ml sterilin tube. The tube is then filled to the 10 ml mark with sterile glycerol. Mix by gently shaking the tube. The volume marks on the sterilin tube are used as it is not possible to accurately pipette 5 ml of undiluted glycerol.
Recipe for TA buffer
Keywords: TA buffer; ligation; Zappe; electrophoresis
Recipes: TA buffer
Contributor: Zappe, H
Related: Gel based DNA manipulation
TA buffer
Component stock conc final conc. / 10ml
Tris-HCl, pH 8.0 1M
NaAcl 5M
Water ???- 5.58ml
??
Recipe for Tris-borate buffer
Keywords: Zappe; TB; electrophoresis; gel
TB buffer
Contributor: Zappe, H
Related: Gel electrophoresis
Tris-borate buffer (5x)
Component stock conc final conc. / 1000ml
Tris-base 54g
Boric acid 27.5g
EDTA, pH 8.0 0.5M 20ml
Do not store Tris-borate in a glass bottle.
Recipe for TE Buffer
Keywords: TE buffer; Babb
TE buffer
Contributor: Zappe, H
Related: many protocols
TE Buffer
Component stock conc final conc. / 10 ml /100ml /1l
Tris, pH 8.0 1M 10mM 0.1ml 1ml
10ml
EDTA, pH 8.0 0.5M 1mM 20ml 0.2ml
2ml ??
Recipe for calcium chloride
Keywords: Zappe; calcium chloride; cacl2
Calcium chloride
Contributor: Zappe, H
Related: many protocols
Storage: 4 C
Calcium chloride (1M), CaCl2
Component stock conc final conc. / 100 ml /1l
CaCl2.2H20 - 1M 14.7g
water - - to 100ml
autoclave
Recipe for chloroform:isoamyl alcohol mixture
Keywords: chloroform:isoamyl alcohol mix; Babb
Chloroform:isoamyl alcohol mix
Contributor: Zappe, H
Related: many protocols
Care:
caution
Chloroform:Isoamyl (24:1)
Component stock conc final conc. / 10 ml /100ml
Chloroform - - - 96ml
Isoamyl alcohol - - - 4ml
Recipe for cysteine
Keywords: cysteine
Recipes: Cysteine
Contributor: Zappe, H
Related: -
Cysteine
Component stock conc final conc. / 10 ml
Cysteine 0.1M 1mM 0.1ml
Water - to 4.69ml
DNA digestion: restriction enzyme dilution buffer
Keywords: Zappe; DNA; digestion; restriction
Restriction enzyme dilution buffer
Contributor: Zappe, H
Related: DNA digestion
Storage: -20 C
Restriction enzyme dilution buffer
Component stock conc final conc. / 10ml
Tris-HCl, pH 7.5 1M 10mM 0.1ml
NaCl 5M 50mM 0.1ml
b mercapto-ethanol 10mM 7 ml
Gelatin 10mg/ml 100 mg/ml 0.1ml
water 5.3ml
glycerol 44%(v/v) 4.4 ml
DNA digestion: restriction enzyme core reaction buffer
Keywords: Zappe; DNA; digestion; restriction
Restriction enzyme core buffers (10X).
Contributor: Zappe, H
Related: DNA digestion
Storage: -20 C
Restriction enzyme dilution buffer (10x)
Salt concentration mM
Component stock conc 0 50 100 150
Tris-HCl, pH 7.5 1M 1ml 1ml 1ml
1ml
NaCl 5M 0 1ml 2ml
87.7mg
BSA (albumin) 10mg/ml 1ml 1ml 1ml 1ml
MgCl2 1M 1ml 1ml 1ml 1ml
DTT 0.5M 0.2ml 0.2ml 0.2ml 0.2ml
water 2.4ml 1.4ml 0.4ml 2.4ml
glycerol 44%(v/v) 4.4 ml 4.4 ml 4.4ml
DNA digestion: Sma I restriction enzyme reaction buffer
Keywords: Zappe; DNA; digestion; restriction; Sma I
Sma I restriction enzyme reaction buffer
Contributor: Zappe, H
Related: DNA digestion
Storage: -20 C
Sma I restriction enzyme reaction buffer (10x)
Component stock conc final conc. / 10ml
Tris-HCl, pH 8.0 1M 0.1M 1ml
KCl 5M 0.2M 2ml
MgCl2 1M 0.2M 2ml
DTT 0.5M 0.2ml 0.2ml
water 0.4ml
glycerol 44%(v/v) 4.4ml
All the components, except the glycerol, are combined in a 10 ml sterilin tube. The tube is then filled to the 10 ml mark with sterile glycerol. Mix by gently shaking the tube. The volume marks on the sterilin tube are used as it is not possible to accurately pipette 5 ml of undiluted glycerol.
Recipe for DNA extraction cell suspension buffer
Keywords: Cell suspension buffer; DNA extraction; Zappe
Cell suspension buffer
Contributor: Zappe, H
Related: Bacterial DNA extraction
Cell suspension buffer:
Component stock conc final conc. / 100 ml /1l /10ml
Tris-HCl (pH 8.0) 1M 10mM 1ml
EDTA (pH 8.0) 0.5M 1mM 0.2ml
sucrose - 0.35M 12g
water - - to 100ml
If the cells are going to be physically disrupted (French press or glass beads), then do not add the sucrose.
Recipe for DNA extraction lysing solution
Keywords: Lysing solution; DNA extraction; Zappe
Lysing solution
Contributor: Zappe, H
Related: Bacterial DNA extraction
Lysing solution (2X):
Component stock conc final conc. / 100 ml /1l /10ml
Tris-HCl (pH 8.0) 1M 100mM 10ml
EDTA (pH 8.0) 0.5M 20mM 4ml
NaCl 5M 0.3M 6ml
SDS 25%(w/v) 2% 8ml
b-mercaptoethanol - 2%(v/v) 2ml
proteinase K 20mg/ml 100mg/ml 0.5ml
water - - to 100ml
Add proteinase K (50 *l of a 20 mg/ml stock solution per 10 ml 2X lysing solution) and the *- mercaptoethanol to the amount of lysing solution needed. The other Components can be premixed as a stock solution.
Recipe for DTT
Keywords: Zappe; DTT; dithiothreitol
Dithiothreitol (DTT)
Contributor: Zappe, H
Related: many protocols
Storage: -20 C
Dithiothreitol (DTT)
Component stock conc final conc. / 20 ml /1l
DTT - 1M 3.09g
sodium acetate, pH 5.2 - - to
20ml
filter sterilize and store aliquots at -20 C
Recipe for EDTA
Keywords: EDTA; ethyl???; Zappe
EDTA (ethyl??)
Contributor: Zappe, H
Related: many protocols
EDTA (Ethylene??)
Component stock conc final conc. / 400 ml /1l
EDTA - 0.5M 93.05g
NaOH - - +/- 10g
water - - to 400ml
EDTA will not dissolve until +/- pH 8.0
autoclave
Recipe for ethidium bromide
Keywords: Zappe; EtBr; ethidium bromide
Ethidium bromide (EtBr)
Contributor: Zappe, H
Related: many protocols
Care:
never breath dust - mutagen
Ethidium bromide (EtBr)
Component stock conc final conc. / 10 ml /1l
EtBr - 10mg/ml 0.1g
water - - to 10ml
Recipe for ethanol stock
Keywords: ethanol; EtOH
Ethanol (EtOH)
Contributor: Zappe, H
Related: many protocols
Ethanol (EtOH)
Component stock conc final conc. / 500 ml /1l
EtOH - 70% 350ml
water - - 150ml
FROZEN COMPETENT&nb