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30% Acrylamide

29g acrylamide
1g of N,N'-methylbisacrylamide
Dissolve in 60ml of water, adjust the volume to 100ml with water.
Filtration through 0.45 micron pore size filter.
Store at 40C.

25mg/ml Ampicillin stock solution recipe

Dissolve 1 g ampicillin (sodium salt) in 35 ml of distilled water.


Make up volume to 40 ml with distilled water.
Sterilize by filtration (0.2µ filter).

0.5 M EDTA (pH 8.0)

Dissolve 186.12g Na2EDTA·2H2O (MW- 372.24) in 800 ml distilled.


Adjust pH to 8.0 with NaOH (~20g of NaOH pellets) [EDTA will dissolve at pH
8.0]
Adjust volume to 1 liter with distilled water.
Sterilize by autoclaving and store at room temperature.

10 mg/ml Ethidium bromide recipe

Dissolve 1g ethidium bromide in 100 ml distilled water. Stored in dark bottle or


wrap in aluminium foil.

Freeze medium

70% basal medium


20% FBS
10% DMSO

6 X Gel loading buffer

Dissolve 25 mg bromophenol blue and 25 mg xylene cyanol FF


Add 3 ml glycerol.
Make up volume to 10 ml with distilled water.
Store at 4°C.
Sachin   Indian Institute of Technology Guwahati/ Sachin/Protocol‐1 
 
Glycerol stock for bacteria storage

65% glycerol
0.1 M MgSO4
0.025 M Tris-HCl pH 8

1M IPTG

1.19g Isopropyl-b,D-thiogalactoside (IPTG, MW 238.3) in 5 ml distilled water.


Sterilize by filtration (0.2µ filter).
Store at -200C.

1x LB medium (Luria-Bertani medium)

Tryptone, 10 g
Yeast extract, 5 g
NaCl, 10 g
Add distilled water to 950 ml
Adjust the pH to 7.0 with 5 N NaOH.
Adjust the volume to 1 liter with distilled water.
Sterilize by autoclaving.

10M NaOH

40g NaOH in 100ml H2O.

1X Phosphate buffer saline

Dissolve the following in 800ml distilled H2O.


8g of NaCl
0.2g of KCl
1.44g of Na2HPO4
0.24g of KH2PO4
Adjust pH to 7.4.
Adjust volume to 1L with additional distilled water.
Sterilize by autoclaving.

Sachin   Indian Institute of Technology Guwahati/ Sachin/Protocol‐1 
 
25:24:1 Phenol:Chloroform:Isoamyl-alcohol

25ml Phenol,
24ml Chloroform,
1ml Isoamyl alcohol.

10 mM Phenylmethylsulfonyl fluoride (PMSF)

Dissolve 17.4 mg PMSF in 10 ml isopropanol.


Aliquot and store at -20°C.

20% SDS

200 g electrophoresis-grade SDS in 900 ml of water.


Heat to 68°C and stir to assist dissolution.
Adjust the volume to 1 liter with water.
Store at room temperature.
Do not autoclave.

1XSOC

Tryptone, 20 g
Yeast extract, 5 g
NaCl, 0.5 g
add water to 950 ml

3M sodium acetate

408 g sodium acetate.3H2O (MW 136).


Dissolve in 800 ml of H2O
Adjust the pH to 5.2 with glacial acetic acid.
Adjust to 1 liter with ddH2O
Sterilize by autoclaving

1X TAE (TRIS/ACETATE/EDTA) buffer

Sachin   Indian Institute of Technology Guwahati/ Sachin/Protocol‐1 
 
242 g of Tris base
57.1 ml of glacial acetic acid
100 ml of 0.5 M EDTA (pH 8.0)
in 1000ml water.

12.5mg/ml Tetracycline solution

Dissolve 1 g tetracycline (yellow powder) in 75 ml of 1:1 vol/vol distilled


water:ethanol.
Make up volume to 80 ml with 1:1 vol/vol distilled water:ethanol.
Sterilize by filtration (0.2µ filter)
Wrap aliquots in aluminium foil and stored at -20 °C.

1M Tris-HCl, pH 6.8

12.1g Tris base (MW 121.14) in 100ml water.


Adjust pH with concentrated HCl.

2% X-gal

Dissolve X-gal in dimethylformamide at a concentration of 20 mg/ml solution.


Store at -20°C and avoid light.

Sachin   Indian Institute of Technology Guwahati/ Sachin/Protocol‐1 
 

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