This is a partially completed html page in a set of molecular biology techniques, recipes and protocols pages. Please read the disclaimer and the introductory pages. Your submissions and comments are always welcome. The pages have been assembled by Brendan Babb at the department of Medical Microbiology, University of Cape Town.

Recipe for S1 mix

Keywords: Zappe; S1 mix; Heinekoff
Recipes: S1 mix

Contributor: Zappe, H

Related: Heinekoff shortening

S1 mix

Component stock conc final conc. / 10 aliquots

S1 buffer, 10x 10x 1x 27 ml

water 172 ml

S1 nuclease 60U- 1 ml

Recipe for S1 stop

Keywords: Zappe; S1 stop; ExoIII; shortening
Recipes: S1 stop

Contributor: Zappe, H

Related: Heinekoff shortening

S1 stop

Component 	stock conc	final conc.	/ 100 ml	/1l	/10ml
Tris base		0.3M
EDTA		10mM
Water		-	to 100 ml



 
Recipe for Sodium dodecyl sulphate 

Keywords: SDS; sodium dodecyl sulphate; Zappe; laural 
sulphate 
   Sodium dodecyl sulphate

Contributor:     
Zappe, H

Related:     
many protocols

Care:







sodium dodecyl sulphate, laural sulphate (SDS) (25%)


Component 	stock conc	final conc.	/ 100 ml	/1l
SDS	-	25%(w/v)	-	
water	-	-	to 100ml



heat to about 80 C to dissolve and add a few fdrops of hydrochloric acid to pH 7.2




 
Recipe for acrylamide stock solution 

Keywords: Zappe; acrylamide 
   Acrylamide-bis-acrylamide stock solution

Contributor:     
Zappe, H

Related:     
SDS-PAGE gels

Storage:     
in dark





Acrylamide-bis-acrylamide stock solution


Component 	stock conc	final conc.	/ 100ml
Acrylamide			29.2 g
bis-Acrylamide			0.8 g
water			to 100ml





Filter through Whatman's No1 paper and store in a dark bottle at 4  C




 
Recipe for SDS-PAGE gel destain 

Keywords: Zappe; destain 
   SDS-PAGE gel destain

Contributor:     
Zappe, H

Related:     
SDS-PAGE gels

Storage:     
RT





SDS-PAGE gel destain


Component 	stock conc	final conc.	/ 500ml
Ethanol	100%	20%	
glacial acetic acid		10%	
water			to 500ml

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Recipe for acrylamide gel mixes

Keywords: Zappe; acrylamide; gel
Acrylamide gel mixes

Contributor: Zappe, H

Related: SDS-PAGE gels

Storage: -

Caution;

acrylamide and TEMED are dangerous

Gel Mixtures

Component 	resolving gel	stacking gel	(7.5 %)	(12.5 %)
Resolving gel buffer	1.25 ml	1.25 ml	   --
Stacking gel buffer	    --	     --	1.25 ml
water	2.5 ml	1.65 ml	3.0 ml
Acrylamide Solution	1.25 ml	2.05 ml	750 ml
APS	15 ml	15 ml	15 ml
TEMED	10 ml	10 ml	10 ml

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Recipe for resolving gel buffer

Keywords: Zappe; resolving buffer
Lower (resolving) gel buffer

Contributor: Zappe, H

Related: SDS-PAGE gels

Storage: RT

Lower (resolving) gel buffer

Component 	stock conc	final conc.	/ 200ml
Trizma base		1.5M	36.35g
SDS		0.4%(w/v)	0.8g
water			to 200ml

Adjust to pH 8.8 with concentrated HCl.

Recipe for sample buffer, SDS-PAGE gels

Keywords: Zappe; sample buffer
Sample buffer (SDS-PAGE)

Contributor: Zappe, H

Related: SDS-PAGE gels

Storage: -20 C

Sample treatment buffer

Component 	stock conc	final conc.	/ 100ml
Stacking gel buffer			29.2 g
SDS 	10%		4ml
Glycerol			2ml
2-mercaptoethanol			1ml
water			0.5ml

Recipe for stacking buffer

Keywords: Zappe; stacking buffer
Upper (stacking) gel buffer

Contributor: Zappe, H

Related: SDS-PAGE gels

Storage: RT

Upper (stacking) gel buffer

Component 	stock conc	final conc.	/ 200ml
Trizma base		0.5M	12.1g
SDS		0.4%(w/v)	0.8g
water			to 200ml

Adjust to pH 6.8 with concentrated HCl

Recipe for SDS-PAGE gel stain

Keywords: Zappe; stain
SDS-PAGE gel stain

Contributor: Zappe, H

Related: SDS-PAGE gels

Storage: RT

SDS-PAGE gel stain

Component 	stock conc	final conc.	/ 500ml
Coomassie Brilliant Blue -R250			1.25 g
methanol			227 ml
water			227 ml
Glacial Acetic Acid			46 ml

Dissolve stain in methanol, add water and mix. Finally add acetic acid. Leave to stand overnight and filter. The above volume of stain can be used for about 8 gels.

Recipe for tank buffer (SDS-PAGE)

Keywords: Zappe; tank buffer
Tank buffer

Contributor: Zappe, H

Related: SDS-PAGE gels

Storage: -

Electrophoresis (or tank) buffer (10X)

Component 	stock conc	final conc.	/ 500ml
Trizma base			15.15 g
Glycine			72.05g
SDS			5.0g
water			to 100ml

Recipe for sodium acetate

Keywords: sodium acetate; Zappe; NaAc
Sodium acetate

Contributor: Zappe, H

Related: many protocols

sodium acetate, NaAc (pH 5.2 or 6.0)

Component 	stock conc	final conc.	/ 500 ml	/100ml
sodium acetate	-	3M	204.5g	
water 	-	-	to 400ml
glacial acetic acid	-	-	to required pH
water (after pH adjust)	-	-	to 500ml

make to pH 5.2 or to pH 6.0

an alternative solution III for plasmid preparation.

Recipe for sodium chloride

Keywords: Zappe; NaCl; sodium chloride
Sodium chloride

Contributor: Zappe, H

Related: many protocols

Sodoium chloride, NaCl (5M)

Component 	stock conc	final conc.	/ 100 ml	/1l
NaCl	-	5M	29.22g	
water	-	-	to 100ml

autoclave.

Recipe for sodium hydroxide (10M)

Keywords: Zappe; sodium hydroxide; NaOH
Sodium hydroxide

Contributor: Zappe, H

Related: many protocols

Care:

caustic

Sodium hydroxide, NaOH, (10M)

Component 	stock conc	final conc.	/ 100 ml	/1l
NaOH	-	10M	40g	
water	-	-	to 100ml

store in a plastic container

dilute to make stocks of 0.1M and 1M

Magnesium sulphate

Contributor: Zappe, H

Related: many protocols

Magnesium sulphate, MgSO4 (1M)

Component 	stock conc	final conc.	/ 100 ml	/1l
MgSO4 (.7H20??)	-	1M	24.7g	
water	-	-	to 100ml

autoclave.

Recipe for sodium perchlorate

Keywords: sodium perchlorate; Zappe
Sodium perchlorate

Contributor: Zappe, H

Related: many protocols

Caution:

can be explosive

sodium perchlorate

Component 	stock conc	final conc.	/ 10 ml	/100ml
NaClO4  	-	5M	7g	70g

  • Filter sterilize

    Recipe for hybridization solution (Zappe)

    Keywords: Zappe; Southern; hybridization
    Pre-hyb/hyb solution

    Contributor: Zappe, H

    Related: Southern transfer DNA

    Storage: RT

    Component 	stock conc	final conc.	/ 200ml
		9 ml 20 X SSC
		0.6 ml 25% SDS
		1.2 ml BLOTTO ( Sambrook et al., page B15)
		0.3 ml 10 mg/ml boiled Salmon sperm DNA
		18.9 ml water

water			to 200ml

    Recipe for Southern transfer solution 1

    Keywords: Babb; Southern; hybridization; ST1; depurination
    Depurination solution (ST1)

    Contributor: Babb, B

    Related: Southern transfer DNA

    Storage: RT

    Depurination solution (ST1)

    Component 	stock conc	final conc.	/ 200ml
HCl	conc.	0.25M	??
water			to 200ml

    Recipe for Southern transfer solution 2

    Keywords: Babb; Southern; hybridization; ST2; denaturation
    Denaturnation solution (ST2)

    Contributor: Babb, B

    Related: Southern transfer DNA

    Storage: RT

    Denaturation solution (ST2)

    Component 	stock conc	final conc.	/ 200ml
NaOH	10M	0.5M	
NaCl	5m	1.5M	
water			to 200ml

    Recipe for Southern transfer solution 3

    Keywords: Babb; Southern; hybridization; ST3; neutralizing
    Neutralizing solution (ST3)

    Contributor: Babb, B

    Related: Southern transfer DNA

    Storage: RT

    Neutralizing solution (ST3)

    Component 	stock conc	final conc.	/ 200ml
Ammonium acetate		1M
NaOH		0.02M
water			to 200ml

    Recipe for STE

    Keywords: STE
    STE

    Contributor:

    Related: -

    Storage: RT

    STE

    Component 	stock conc	final conc.	/ 200ml
Tris buffer, pH 8.0	1.0M	10mM	2ml
EDTA, pH 8.0	0.5M	1mM	0.4ml
NaCl	5M	100mM	4ml
water			to 200ml