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2024年12月29日发(作者：适合编程的平板电脑)
Table of Contents
.1
2
3
4
6X 5
10 6
10 7
20 8
1% SDS, 0.2 9
14% PEG (8000), 2M NaCl, 10 .10
20% 11
1.0 M Tris, pH 8.0, 1.5 12
10mM Tris-HCl, pH 7.5, 13
10 mM Tris, 50 mM EDTA, pH 14
10 mM Tris-HCl, 1 mM EDTA, pH 15
3 M Sodium Acetate, pH 16
.17
Labelling 18
Sequencing .19
5% .20
6% Acrylamide in TBE, 50% 21
40% 22
LB Medium (1 Liter)
10g Bacto-tryptone
5g Bacto-yeast extract
10g NaCl
For forty plates add 1% agar--1g. Autoclave media. When cool, add ampicillin
and pour plates. For 1L of media, add 1.8 mL amp.
NZ Medium (500 mL)
5 g Bacto-tryptone
2.5 g Bacto-yeast extract
2.5 g NaCl
1.25 g MgSO4
For 20 plates add 1.2% agar--6g. Autoclave and pour plates at 50o C
SM Buffer (1L)
5.8 g NaCl
1.2 g MgSo4
50 mL 1M Tris-HCl, pH 7.5
0.1 g Gelatin (doesn't dissolve)
Autoclave
Used for phage dilution and storage.
SET Buffer
50 mM Tris-HCl, pH 8.0, 50 mM EDTA, 20% w/v Sucrose
to make 200mL:
40 g Sucrose
10 mL of 1M Tris
20 mL of 0.5 M EDTA, disodium salt
bring to 200 mL with H20
6X Prehybridization Solution
to make 500 mL
300 mL ddH20
150 mL 20X SSC
50 mL 50X Denhardt's solution
1 mL 0.5 M EDTA (disodium salt)
2.5 mL 20% SDS
6X refers to the concentration of SSC
10X TBE Buffer (for polyacrylamide gels)
to make one liter:
60.75 g Tris
3.7 g EDTA (tetrasodium salt)
30 g Boric acid
10X TAE Buffer (For agarose gels)
to make one liter:
48.20 g Tris
6.75 g NaAce
3.75 g EDTA (disodium salt)
Adjust pH to 7.6 with acetic acid. (Approx. 20 mL)
20X SSC
to make one liter:
175.3 g NaCl
88.2 g NaCitrate
add water to bring volume to one liter.
adjust to pH 7.0 with HCl.
1% SDS, 0.2 M NaOH
to make 100 mL:
93 mL ddH20
5 mL 20% SDS
2 mL 10 M NaOH
14% PEG (8000), 2M NaCl, 10 mM MgSO4
to make one liter:
140 g PEG
117 g NaCl
2.46 g MgSO4
For use in phage DNA preparation.
20% SDS
to male 250 mL:
50 g of SDS in a beaker
Add stir bar and H20 last.
This solution will have to be heated for the SDS to dissolve.
1.0 M Tris, pH 8.0, 1.5 M NaCl
to make one liter:
121.1 g Trizma
87.6 g NaCl
in a volume of water less than 1L. Adjust pH with HCl, then bring to 1L with
H20
10 mM Tris-HCl, pH 7.5, 10 mM MgSO4
to make one liter:
10 mL 1 M Tris-HCl
2.46 g MgSO4
for use in phage DNA preparation
10 mM Tris, 50 mM EDTA, pH 7.5
to make 200 mL:
2 mL 1 M Tris
20 mL 0.5 M EDTA (tetrasodium salt)
178 mL ddH20
adjust pH with HCl.
10 mM Tris-HCl, 1 mM EDTA, pH 7.5
to make 200 mL:
2.0 mL 1 M Tris-HCl, pH 7.5
0.4 mL 0.5 M EDTA
197.6 mL ddH20
3 M Sodium Acetate, pH 4.8
to make one liter:
408.1 g NaAce (trihydrate; gets cold in soln)
about 700 mL H20
adjust pH with glacial acetic acid (takes a lot)
Measure tru pH by dilution with water; range will be between 4.8 and 5.5.
Electrophoresis Dye
to make 4 mL:
3 mL 50 mM EDTA, 10 mM Tris-HCl, pH 8.0
1 mL glycerol
20 L BPB
10 L Xylene cyanol
Stop dye for labelled probe
about 200 l glycerol
add a few grains of blue dextran (8000)
Sequencing gel dye
for approx 1 mL:
1 mL formamide
10 L xylene cyanol
10 l BPB
3 L 10 M NaOH
5% acrylamide
to make 200 mL:
20 mL 10X TBE
25 mL 40% acrylamide
155 mL H20
6% Acrylamide in TBE, 50% Urea
to make 500 mL:
50 mL 10X TBE
75 mL 40% acrylamide
250 g Urea
bring to 500 mL with H2O
40% Acrylamide (38:2 acrylamide:bis acrylamide)
to make 200 mL:
76 g acrylamide
4 g bis acrylamide
bring to 200 mL with H2O
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