1. .1% agarose with EB
Melt 10g of agarose in 1 liter of 1xTBE and add 20ul of EB(10mg/ml), then store at 65c water bath .You can dispense
according to your requirement.
2. 10M Ammonium Acetate(50ml)
Ammonium acetate
38.5g
H2O
to 50ml
3. B broth(per liter)
Bacto-tryptone
10g
NaCl
8g
H2O
to
1 liter
After autoclaving and cooling add 10ml of sterile 20% glucose.
4. B top agar
B broth plus 6g Bacto-agar per liter , after autoclaving and cooling , add 10ml of sterile 20% glucose.
5. B plates
B broth plus 15g Bacto-agar per liter. After autoclaving and cooling
add 10ml of sterile 20% glucose.
6. Bromo phenol blue
10mg/ml dissolve in SDW
7. 2 x CTAB buffer(for plant DNA preps)
2% CTAB (w/v)
20g
100mM Tris(PH 8.0 , 1M)
100ml
20mM EDTA(PH 8.0, 0.5 M)
40ml
1.4 M NaCl
81.8g
1%PVP(Mr40,000)
5g
d H2O
to 1000ml
Autoclave
8. 10%CTAB solution
10% CTAB
100g/ 1 liter
0.7 M NaCl
41g/ 1 liter
d H2O
to 1000ml
Autoclave
9. CTAB precipitation buffer
1% CTAB
10g
50mM Tris(PH8.0, 1 M)
50ml
10mM EDTA(PH8.0, 0.5 M)
20ml
d H2O
to 1000ml
Autoclave
10. CTAB buffer (for mini preps of plant genomic DNA and purification of
large scale genomic DNA)
0.2M Tris-HCl(PH7.5, 1M)
200ml
0.05M EDTA(PH8.0, 0.5M)
100ml
2.0M NaCl
116.7g
2%W/V CTAB
20g
d H2O
to 1000ml
Autoclave
11. CTAB solution(for antirrhinum DNA preps)
50mM Tris PH 7.45(12.5ml 1M Tris)
10mM EDTA PH8.0(5ml 0.5M EDTA)
2% CTAB(5.0 g)
H2O to 250ml
12. 50 x Denhardt’s
10 g
ficoll(Type 400)
10 g
polyvingpyrrolidone(MW.360,000)
10 g
B S A(Fraction v)
H2O
to 1000ml
13.DNA miniprep extraction buffer(Antirrhinum leaves)
0.5M EDTA (PH8.0)
10ml(final
50mM)
5MNaCl
2 ml (final
0.1M)
1 MTris(PH8.0)
10ml(final
0.1M)
10%SDS
10ml(final
1%)
H2O
to
100ml
14.1M Dithiothnetol(DTT) 10ml
DTT
1.55g
10mM sodium acetate(PH 5.2) to 10ml
Store at –20c
15.0.5M EDTA (PH8.0)
EDTA 2H2O
186.1g
H2O
800ml
Adjust PH to 8.0 with NaOH(about 20g of NaOH was needed) and fix the volume to 1000ml, autoclave.
16.10mg/ml Ethidium Bromide(20ml)
Ethidium bromide
H2O
0.2g
to 20ml
Mix well, store at 4c in dark. Handle with extreme caution
17.Formaldehyde gel(100ml)
Agrose
1-1.5g
10 x MOPS buffer
10ml
water
73ml
Dissolve agrose and cool to 50c. Add 17ml of formaldehyde (37%v/v sulotion).
Mix and pour immediately.
Running buffer is 1x MOPs buffer.
18.X-gal
20mg/ml in dimethylformamide(DMF) use a glass or polypropylene tube,wrap in aluminum foil and store at –20c.
19.Hybridization solution( for southern and northern) 1 liter
20x SSC
300ml( final 6x SSC)
50x Denhandt’s
100ml( final 5x Denhardt’s)
10% SDS
50ml( final 0.5%)
SDW
to 1000ml
10mg/ml of ssDNA added to the final 20ug/ml
20.IPTG
2g of IPTG dissolve in 100ml of H2O, Sterilize by filtration, store at –20c at 1ml aliquots.
21.5M KAc(pH~4.8)
461g KAc in H2O, adjust PH with glacial acetic acid and autoclave.
22. LB medium-1 liter
bacto-trypton
10g
bacto yeast extract
5g
NaCl
10g
H2O
950ml
Adjust PH to 7.0 with 5N NaOH and fix the volume to 1000ml, autoclave.
For LB plates, add 15g of bacto-agar to 1 liter medium, autoclave;
For LB top agar, add 7g of bacto-agar per liter LB medium.
23.RNA gel loading buffer(100ml)
Glycerol
50ml
Bromophenol blue
10mg
SDW
to 100ml
24.1M MgSO4
MgSO4 7H2O 246.47g + H2O to 1000ml
25.10 x MOPS
MOPS
46.2g (final 0.2M)
EDTA
3.72g (final 0.01M)
NaAc
41g (final 0.5M)
SDW
800ml
Adjust to 7.0 with HAc and fix the volume to 1000ml with SDW.
26.1M MgCL2
MgCL2 6H2O 20.3g +
H2O
to 100ml
26.10 x phosphate buffer saline (PBS)
NaCL
80g
KCL
2g
Na2HPO4 7H2O
26.8g
KH2PO4
2.4g
H2O
800ml
Adjust the PH to 7.4 with HCL and fix the volume to 1000ml with H2O.
27.10 x PCR buffer(10ml)
1M Tris-HCL(PH 8.3)
1ml
(final 100mM)
1M KCL
5ml
(final 500mM)
1M MgCL2
0.2ml
(final 20mM)
1% Gelatin
1ml
(final 0.1% w/v)
SDW
to
10ml
Make the solution to 10ml by adding SDW(no need for autoclaving)
28.RNase stock solution
10mg/ml of RNase A in H2O.
Heat at boiling(in a water bath) for at least 10 min to destroy any DNase and then kept frozen until needed.
29.3M Sodium acetate
408.1g of NaAc 3H2O in 800ml of H2O, adjust the PH to 5.2 with glacial acetic acid, fix the volume to 1 liter with H2O.
Autoclave.
30.20 x SSC
NaCl
175.3 g
Sodium citrate
88.2 g
H2O
800 ml
PH to 7.0 with a few drops of 10N of NaOH.
Fix the volume to 1 liter and autoclave.
31.5M NaCl
292.2 g of NaCl dissolve in 800ml of H2O and fix the volume to 1000ml, autoclave.
32.10% SDS
100g of SDS in 900ml of H20, heat the solution to 68c to helping dissolve, adjust the PH to 7.2 by adding a few drops of
concentrated HCl(there is no need to sterilize).
33.SOB medium per liter
Trypton
20g
Yeast extract
5g
NaCl
0.5g
250mM KCl
10ml
Add water to 900ml. Adjust PH to 7.0 and add water to 990ml. Autoclave, cool to room temperature and add 10ml of sterile
solution of 1M MgCl2 before use.
34.SOC medium per liter
SOB with the addition of 20ml filter sterilized 1M glucose per liter of SOB.
35.1M Tris
121.1 g
Tris BASE
H2O
800ML
Adjust the PH with HCl and fix the volume to 1000ml. Autoclave.
36.1 x TE
Tris-HCl (PH 8.0)
10mM
EDTA (PH 8.0)
1mM
37.50 x TAE Buffer/liter
Tris base
242g
Glacial acetic acid
57.1ml
0.5M EDTA,PH8.0
100ml
H2O
to 1 liter
Adjust PH to 8.5.
38.10x TBE buffer/liter
Tris base
108 g
Boric acid
55 g
0.5M EDTA,PH8.0
40ml
H2O
to
1 liter
39.2x YT
bacto-tryptone
16 g
bacto-yeast extract
10 g
NaCl
5g
Adjust PH to 7.0 with 5N of NaOH and fix the volume to 1 liter. Autoclave.