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Reference Guide
Welcome to the Little Book of
Standards: DNA Standards
This handbook is a practical guide to Bio-Rad’s complete line of DNA and protein
standards for electrophoresis and blotting. Bio-Rad standards are an excellent
means of monitoring electrophoresis and blotting experiments, or determining DNA
and protein sizes.
This handbook is organized into five sections for both DNA and protein standards.
On this side of the booklet, DNA standards are divided into sections based on
logical groupings of size ranges in base pairs (bp). On the other side, protein
standards are organized by general application.
Each page in this reference guide offers specifications for each set of standards,
recommended applications, a photograph of the standards run on a gel, and the
length in bp or the mass of each standard band. Each section divider contains
gel migration summaries for the standards described in that section.
We hope you find this booklet to be a useful guide to the selection and use of
Bio-Rad DNA and protein standards. For more information on products or
applications, contact your local Bio-Rad representative, or visit us on the Web
at discover.bio-rad.com
1
Bio-Rad DNA Standards and
Molecular Mass Markers
20 bp–2 kb DNA Standards 3–8 5 kb–5.7 Mb DNA Standards 22–28
20 bp Molecular Ruler 4 CHEF DNA Size Standards, 5 kb Ladder 23
EZ Load™ 20 bp Molecular Ruler 5 CHEF DNA Size Standards, 8 – 48 kb Ladder 24
AmpliSize® Molecular Ruler 6 CHEF DNA Size Standards, Lambda Ladder 25
100 bp Molecular Ruler 7 CHEF DNA Size Markers, 26
EZ Load 100 bp Molecular Ruler 8 S. cerevisiae Chromosomes
CHEF DNA Size Markers, 27
100 bp–10 kb DNA Standards 9 –17 H. wingei Chromosomes
EZ Load™ HT 100 bp –2 kb Molecular Marker 10 CHEF DNA Size Markers, 28
EZ Load HT 500 bp–10 kb Molecular Marker 11 S. pombe Chromosomes
100 bp PCR Molecular Ruler 12
Fluorescent DNA Standards 29–31
EZ Load 100 bp PCR Molecular Ruler 13 100 bp Fluorescein-Labeled Ruler 30
500 bp Molecular Ruler 14 100 bp Texas Red-Labeled Ruler 31
EZ Load 500 bp Molecular Ruler 15
Precision Molecular Mass Ruler 16
EZ Load Precision Molecular Mass Ruler 17
2
20 bp–2 kb DNA Standards
Bio-Rad molecular rulers are available in several convenient size ranges and
increments for sizing single- and double-stranded DNA. Visually distinct reference
bands make it easy to determine the size of your sample DNA.
2,000 bp
1,500
1 kb
800 bp 800 bp
700
500 500 500
100 bp
60
50
40
20
10 bp
20 bp 100 bp AmpliSize®
Molecular Molecular Molecular
DNA Standards
Ruler Ruler Ruler
20 bp–2 kb
3
20 bp Molecular Ruler
Size Range Quantity Recommended Load Volume
60
40
20
4
EZ Load™ 20 bp Molecular Ruler
Size Range Quantity Recommended Load Volume
20
5
®
AmpliSize Molecular Ruler
Size Range Quantity Recommended Load Volume
polyacrylamide. 400
100
50
6
100 bp Molecular Ruler
Size Range Quantity Recommended Load Volume
Catalog #: 170-8202
100
7
EZ Load™ 100 bp Molecular Ruler
Size Range Quantity Recommended Load Volume
Catalog #: 170-8352
100
8
100 bp–10 kb DNA Standards
Bio-Rad molecular rulers are available in several convenient size ranges and
increments for sizing single- and double-stranded DNA. Visually distinct reference
bands make it easy to determine the size of your sample DNA.
10 kb
10,000 bp
8,000 bp
5,000
4,000
3,000 bp
1 kb
1,000 1,000 1,000 bp (100 ng)
700 (70 ng)
500 500 500 500 500 (50 ng)
DNA Standards
400
100 bp– 10 kb
300
100 bp
100 100 bp (10 ng)
9
EZ Load™ HT 100 bp–2 kb
Molecular Marker
Size Range Quantity Recommended Load Volume
Catalog #: 170-8361
10
EZ Load™ HT 500 bp–10 kb
Molecular Marker
Size Range Quantity Recommended Load Volume
500
Recommended gels: 1% ReadyAgarose 96 Plus
precast gel; 0.8 –1.5% standard agarose.
Catalog #: 170-8362
11
100 bp PCR Molecular Ruler
Size Range Quantity Recommended Load Volume
Catalog #: 170-8206
500
100
12
EZ Load™ 100 bp
PCR Molecular Ruler
Size Range Quantity Recommended Load Volume
100
13
500 bp Molecular Ruler
Size Range Quantity Recommended Load Volume
Catalog #: 170-8203
1,000
500
14
EZ Load™ 500 bp Molecular Ruler
Size Range Quantity Recommended Load Volume
Catalog #: 170-8354
1,000
500
15
Precision Molecular Mass Ruler
Size Range Quantity Recommended Load Volume
Catalog #: 170-8207
16
EZ Load™ Precision
Molecular Mass Ruler
Size Range Quantity Recommended Load Volume
Catalog #: 170-8356
17
1–35 kb DNA Standards
Bio-Rad molecular rulers are available in several convenient size ranges and
increments for sizing single- and double-stranded DNA. Visually distinct reference
bands make it easy to determine the size of your sample DNA.
35.0 kb
30.0
DNA Standards
25.0
1– 35 kb
20.0
15.0 kb 15.0
10 kb 12.5
7.0 7.5
6.0
5.0 5.0
4.0
3.0
2.5
2.0
1 kb
1 kb 2.5 kb
Molecular Molecular
Ruler Ruler
18
1 kb Molecular Ruler
Size Range Quantity Recommended Load Volume
Catalog #: 170-8204
3
19
EZ Load™ 1 kb Molecular Ruler
Size Range Quantity Recommended Load Volume
20
2.5 kb Molecular Ruler
Size Range Quantity Recommended Load Volume
Catalog #: 170-8205
2.5
21
5 kb–5.7 Mb (Pulsed Field)
DNA Standards
DNA Standards
Bio-Rad offers several standards for DNA separations on pulsed field gels,
5 kb– 5.7 Mb
ranging from cosmid inserts to whole chromosomes.
5.7 Mb
4.6
3.13 Mb 3.5
2,200 kb
1,600 2.70
2.35
1.81
1.66
1,125 1.37
1 Mb 1,020 1.05
945
825
785
750
680
610
565
450
365
285
225
100 kb 145.5 kb
97.0
48.5
48.5 kb
38.4
33.5
29.9
24.8
22.6
19.4
17.1
14.7 kb 15.0
10 kb 12.2
10.0
9.8 8.6
8.3
4.9
Catalog #: 170-3624
14.7 kb
9.8
4.9
23
CHEF DNA Size Standards,
8–48 kb Ladder
Size Range Quantity Recommended Load Volume
19.4
17.1
15.0
12.2
10.0
8.6
8.3
24
CHEF DNA Size Standards,
Lambda Ladder
Size Range Quantity Recommended Load Volume
Note: Cut a plug from a block into a size that will fit
a well of your gel (5 – 8 plugs per block). The height
of the plug should not extend above the well. After
placing the plug into a well, seal the well with melted
1.0% low-melt agarose and allow it to solidify. Store
blocks at 4°C.
Catalog #: 170-3635
145.5 kb
97
48.5
25
CHEF DNA Size Markers,
S. cerevisiae Chromosomes
Size Range Contents Recommended Load Volume
Note: Cut a plug from a block into a size that will fit 1.13
a well of your gel (5– 8 plugs per block). The height 1.0
0.95
of the plug should not extend above the well. After
0.83
placing the plug into a well, seal the well with melted 0.79
1.0% low-melt agarose and allow it to solidify. Store 0.75
26
CHEF DNA Size Markers,
H. wingei Chromosomes
Size Range Contents Recommended Load Volume
Note: Cut a plug from a block into a size that will fit
a well of your gel (5– 8 plugs per block). The height of
the plug should not extend above the well. After
placing the plug into a well, seal the well with melted
1.0% low-melt agarose and allow it to solidify. Store
2.70
blocks at 4°C.
2.35
Catalog #: 170-3667
1.81
1.66
1.37
1.05
27
CHEF DNA Size Markers,
S. pombe Chromosomes
Size Range Contents Recommended Load Volume
Catalog #: 170-3633
28
DNA Standards
Fluorescent
Fluorescent DNA Standards
Bio-Rad fluorescent DNA standards are ideal for fast, accurate imaging.
A choice of two fluorescent dyes provides multiplexing capabilities.
10 kb
1 kb
800 bp 800 bp
500 500
400 400
300 300
200 200
100 bp
100 bp 100 bp
Fluorescein- Texas Red-
Labeled Labeled
Ruler Ruler
29
100 bp Fluorescein-Labeled Ruler
Size Range Quantity Recommended Load Volume
Catalog #: 170-8216
30
100 bp Texas Red-Labeled Ruler
Size Range Quantity Recommended Load Volume
Catalog #: 170-8217
31
Texas Red is a trademark of Molecular Probes, Inc.
Bio-Rad
Laboratories, Inc.
Life Science Web site www.bio-rad.com USA (800) 4BIORAD Australia 02 9914 2800
Austria (01)-877 89 01 Belgium 09-385 55 11 Brazil 55 21 507 6191
Group Canada (905) 712-2771 China (86-21) 63052255
Czech Republic +420 2 41 43 05 32 Denmark 44 52 10 00
Finland 09 804 22 00 France 01 47 95 69 65 Germany 089 318 84-177
Hong Kong 852-2789-3300 India (91-124)-6398112/113/114, 6450092/93
Israel 03 951 4127 Italy 39 02 216091 Japan 03-5811-6270
Korea 82-2-3473-4460 Latin America 305-894-5950
Mexico 55-52-00-05-20 The Netherlands 0318-540666
New Zealand 64 9 415 2280 Norway 23 38 41 30 Poland +48 22 331 99 99
Portugal 351-21-472-7700 Russia 7 095 721 1404
Singapore 65-6415 3188 South Africa 00 27 11 4428508
Spain 34 91 590 5200 Sweden 08 555 12700 Switzerland 061 717-9555
Taiwan (8862) 2578-7189/2578-7241 United Kingdom 020 8328 2000
This handbook is organized into five sections for both protein and DNA standards.
On this side of the booklet, protein standards are divided into sections based
on applications. Featured are our new Precision Plus Protein™ standards,
highly purified recombinant proteins that offer a new level of band clarity and
application versatility.
Each page in this reference guide offers specifications for each set of standards,
recommended applications, a photograph of the standards run on a gel, and the
molecular mass or pI of each standard band. Each section divider contains gel
migration summaries for the sets of standards described in that section.
We hope you find this booklet to be a useful guide to the selection and use
of Bio-Rad protein and DNA standards. For more information on products or
applications, contact your local Bio-Rad representative, or visit us on the Web
at discover.bio-rad.com
1
Bio-Rad Protein Molecular Weight
and Reference Standards
Precision Plus Protein™ Standards 3 –7 Blotting Standards 22– 25
Precision Plus Protein Standards, Unstained 5 Biotinylated SDS-PAGE Standards, 23
Precision Plus Protein Standards, All Blue 6 Broad Range
2
Precision Plus Protein™ Standards
Protein Migration Chart
250
250 150 250
250 150 100
250 100 75 250 250 150
150 150
100 75 50 250 100 150 100
250 150 250
37 100
75 75
50 150 75 75
100
150 150 50
250 37 25 100 50
75 50 37 50
20 100 75 37
100
37
75 50 37
25 15 25
50 37 25
150 75 20 50 20 20
10 25
37 25 37 15 25
20
15
20 15 15 10 20
100 50 25
20 10
10 15
15 10 15
37 10 10 10
Protein Standards
75 25
Precision Plus
Tris-HCI Gels
Protein molecular mass, kD. Migrations based on running gels until the dye front reached the bottom
of the gel.
3
Precision Plus Protein™ Standards
Protein Migration Chart
250
250 250 250 150
150 150 250 250 250
150 100
100 100
150 150
75 75 150 250
75 100
50 100
50 75 75 50 100 100
37 150
37 75
50 37
25 75
37 50 100
25 20 50
20 15 25 37 75
37 25
15 20 50
20
10 15
25 50
10 20 37
10 25 15
37
20 15
10
Protein molecular mass, kD. Migrations based on running gels until the dye front reached
the bottom of the gel.
4
Precision Plus Protein™ Standards,
Unstained
Size Range Quantity Recommended Load Volume
10
* Allow standards to reach room temperature, and mix thoroughly Stained with
to dissolve any precipitated solids. For certain applications, Coomassie
dilution of the standards is recommended. See instruction R-250.
manual for details.
5
Precision Plus Protein™ Standards,
All Blue
Size Range Quantity Recommended Load Volume
Store at –20°C. 20
15
Catalog #: 161-0373
10
* Allow standards to reach room temperature, and mix thoroughly
to dissolve any precipitated solids. For certain applications,
dilution of the standards is recommended. See instruction
manual for details.
6
Precision Plus Protein™ Standards,
Dual Color
Size Range Quantity Recommended Load Volume
10
7
Unstained Standards
Protein Migration Chart
200 200
200 116 200
116 97.4 116
200 97.4 66
200 116 97.4
66 45 116
200 97.4 200 97.4
200 66
116
66 31 66
97.4 45
116 45
200 116 97.4 45
66
45 116
31 66
97.4 21.5 97.4 31
31
66 45
Standards
Unstained
66 45 31 21.5 14.4
31 21.5 21.5
116 45
Tris-HCI Gels
Protein molecular mass, kD. Migrations based on running gels until the dye front reached
the bottom of the gel.
8
SDS-PAGE Standards,
Broad Range
Size Range Quantity Recommended Load Volume
Catalog #: 161-0317
9
SDS-PAGE Standards,
Low Range
Size Range Quantity Recommended Load Volume
11
Polypeptide SDS-PAGE Standards
Size Range Quantity Recommended Load Volume
-Lactalbumin
Recommended gel: 16.5% Tris-Tricine or 14.4
Catalog #: 161-0326
12
Silver Stain SDS-PAGE Standards,
Low Range
Size Range Quantity Recommended Load Volume
13
Silver Stain SDS-PAGE Standards,
High Range
Size Range Quantity Recommended Load Volume
Catalog #: 161-0315
Ovalbumin 45
Stained
with Bio-Rad
silver stain.
14
SDS-PAGE Standards for
SYPRO Orange, Broad Range
Size Range Quantity Recommended Load Volume
15
Prestained Standards
Protein Migration Chart
5% 7.5% 10% 12% 15% 18% 4–15% 4–20% 8–16% 10–20% 8–16% 10–20%
200
Prestained
200
Standards
200 116 200
116 97.4 116
200 97.4 66
200 116 97.4
66 45 116 26.6
200 97.4 200 97.4
200 66
116
66 31 66 16.9
97.4 45 26.6
116 45 14.4
200 116 97.4 45
66
45 116 16.9
31 66
97.4 21.5 97.4 31
31
66 45
14.4
66 45 31 21.5 14.4
31 21.5 21.5
116 45
6.5
14.4 21.5 14.4 14.4
6.5
97.4 31 21.5 31 14.4 6.5
6.5
45 6.5 6.5 6.5
21.5 3.4
3.4
66 14.4 14.4 1.4 1.4
6.5
16
Prestained SDS-PAGE Standards,
Broad Range
Size Range Quantity Recommended Load Volume
Molecular weights
are representative of
this particular lot.
17
Prestained SDS-PAGE Standards,
Low Range
Size Range Quantity Recommended Load Volume
Molecular weights
are representative
of this particular lot.
18
Prestained SDS-PAGE Standards,
High Range
Size Range Quantity Recommended Load Volume
19
Kaleidoscope™ Prestained Standards
Size Range Quantity Recommended Load Volume
20
Kaleidoscope™ Polypeptide
Standards
Size Range Quantity Recommended Load Volume
Catalog #: 161-0325
21
Blotting Standards
Protein Migration Chart
Standards
Blotting
pH 8.8 gels. Select the acrylamide percentage that best resolves your protein or
peptide of interest.
200 200
200 116 200
116 97.4 116
200 97.4 66
200 116 97.4
66 45 116
200 97.4 200 97.4
200 66
116
66 31 66
97.4 45
116 45
200 116 97.4 45
66
45 116
31 66
97.4 21.5 97.4 31
31
66 45
66 45 31 21.5 14.4
31 21.5 21.5
116 45
Tris-HCI Gels
22
Biotinylated SDS-PAGE Standards,
Broad Range
Size Range Quantity Recommended Load Volume
23
Biotinylated SDS-PAGE Standards,
Low Range
Size Range Quantity Recommended Load Volume
Catalog #: 161-0306
24
Biotinylated SDS-PAGE Standards,
High Range
Size Range Quantity Recommended Load Volume
Catalog #: 161-0311
25
2-D and IEF
Standards
2-D and IEF Standards
Protein Migration Chart
pH 3–10 pH 5–8
8.0 8.0
7.8
7.8
7.5
7.1
7.5
7.0
7.0
6.5
6.5
6.0 6.0
5.1
5.1
4.6
4.6
IEF Gels
26
2-D SDS-PAGE Standards
Range Quantity Recommended Load Volume
27
IEF Standards
Size Range Quantity Recommended Load Volume
28
Protein Gel Stains
Gel Stain Sensitivity* Time Advantages
29
Molecular Weight Determination
Molecular weights of proteins are determined by comparison of their mobilities with
those of several marker proteins (standards) of known molecular weight:
1. After a gel has been run, but before it has been stained, mark the position of
the tracking dye.
2. After staining, measure the migration distance of each protein (standards and
unknowns) from the top of the resolving gel.
a standard curve.
Rf
30
Coomassie is a trademark of Imperial Chemical Industries PLC. Strep-tag and StrepTactin are
trademarks of Institut für Bioanalytik GmbH. SYPRO is a trademark of Molecular Probes, Inc.
Bio-Rad
Laboratories, Inc.
Life Science Web site www.bio-rad.com USA (800) 4BIORAD Australia 02 9914 2800
Austria (01)-877 89 01 Belgium 09-385 55 11 Brazil 55 21 507 6191
Group Canada (905) 712-2771 China (86-21) 63052255
Czech Republic +420 2 41 43 05 32 Denmark 44 52 10 00
Finland 09 804 22 00 France 01 47 95 69 65 Germany 089 318 84-177
Hong Kong 852-2789-3300 India (91-124)-6398112/113/114, 6450092/93
Israel 03 951 4127 Italy 39 02 216091 Japan 03-5811-6270
Korea 82-2-3473-4460 Latin America 305-894-5950
Mexico 55-52-00-05-20 The Netherlands 0318-540666
New Zealand 64 9 415 2280 Norway 23 38 41 30 Poland +48 22 331 99 99
Portugal 351-21-472-7700 Russia 7 095 721 1404
Singapore 65-6415 3188 South Africa 00 27 11 4428508
Spain 34 91 590 5200 Sweden 08 555 12700 Switzerland 061 717-9555
Taiwan (8862) 2578-7189/2578-7241 United Kingdom 020 8328 2000