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Electrophoresis Chemicals

Reagent Package-DNA Gels

Contains all reagents needed to separate DNA restriction fragments by electrophoresis on 35 small agarose gels. Includes electrophoresis buffer (Tris-Acetate-EDTA; enough to make 10 liters), agarose (10 g), Loading buffer (5 ml of a 2 X solution), dye for rapid DNA staining and either methylene blue (5 ml of a 1000 X concentrate) or ethidium bromide (1 ml-5mg/ml) as gel stain. The reagent package is also supplied with an instruction sheet for preparation, electrophoresis and staining of gels.

3-1 with methylene blue $67.45
3-2 with ethidium bromide $67.45

Agarose

DNA electrophoresis grade

3-3-10 10 g $22.04
3-3-25 25 g $43.34

Electrophoresis Buffer (Tris-Acetate-EDTA)

3-4 100 ml of a 100 X solution $31.40

Gel-Loading Buffer

(Bromophenol Blue-Glycerol)

3-5  5 ml of a 2 X solution $11.68

Methylene Blue

Methylene Blue

A stain for DNA in agarose gels.

3-6 5 ml of a 1000 X solution  $12.09

Ethidium Bromide

A stain for DNA that requires UV-illumination for detection. Ethidium Bromide is a mutagen and probably a carcinogen. This chemical should be handled with extreme caution and should in our opinion not be used in an open teaching laboratory.

3-7  l ml of a 5 mg/ml solution $7.21

a Restriction Endonucleases

Each enzyme is supplied with its own enzyme reaction buffer concentrate (10 X concentration) which is provided in a separate tube. Enzymes are also supplied with an instruction sheet for restriction nuclease cutting of DNA. One unit is defined as the amount of enzyme required to completely digest 1µg of phage lambda DNA in 60 minutes.

EcoRI

3-8-1 1000 units  $21.61
3-8-5 5000 units $56.46

BamHI

3-9-1 1000 units $28.83
3-9-5 5000 units $71.99

Hind III

3-9A-1 1000 units  $28.83

 

a Electrophoresis Standards

Dye Mixture

A mixture of xylene cyanol, bromophenol blue and orange G that can be used to estimate the sizes of small DNA fragments on an agarose gel. The dyes are dissolved in gel-loading buffer and ready for electrophoresis.

3-10 5 ml of a 1 X solution $ 13.33

DNA Standard I

Lambda Hind III Digest-Phage Lambda DNA was digested with Hind III, yielding 8 fragments: 23130, 9416, 6557, 4361, 2322, 2028, 564, and 125 base pairs. The DNA is dissolved in gel-loading buffer and is ready for electrophoresis.

3-11

100 µg (400µl)- enough for 30-40 gel lanes

 $50.39

DNA Standard II

Lambda EcoRI, Hind III Digest- Phage lambda DNA was digested with EcoRI and Hind III yielding 13 fragments: 21226, 5148, 4973, 4277, 3530, 2027, 1904, 1584, 1330, 983, 83l, 564, and 125 base pairs. The DNA is dissolved in gel-loading buffer and is ready for electrophoresis.

3-12

100µg (400 µl)- enough for 30-40 gel lanes

 $50.39

DNA Standard III

These DNA samples exhibit the nucleosome repeat of 200 base-pairs when electrophoresed on agarose gels (See page 23, experiment 306). Electrophoretic analysis of this DNA can be used to illustrate the structure of chromatin. The DNAs also provide economical standards for small DNA fragments. The DNAs were obtained by digestion of calf thymus nuclei with low (Nucleosome I), medium (Nucleosome II) and high (Nucleosome III) amounts of micrococcal nuclease. Two of the DNA samples are shown in the picture of the gel on the right (lanes 9, 10). The DNAs are dissolved in gel-loading buffer and are ready for electrophoresis.

3-13

Nucleosome I, II and III 600µl each

 $38.88

 

a DNA

DNA preparations are of the highest of purity and suitable for gene cloning experiments and as substrates for restriction endonucleases. DNAs are shipped as aqueous solutions in wet ice and are supplied with an instruction sheet for restriction nuclease digestion. The Figure below shows electrophoretic analysis of our DNAs after digestion by restriction endonucleases.

   1   2   3   4   5    6   7  8   9   10

The indicated DNA digests were electrophoresed on 1.1% (left panel) or 1.5% (right panel) agarose gels using the PROCELL, PROCELL conversion kit and MB- 170 Power Supply.

 

 

 

 

 

 

 

Lane DNA Restriction Endonuclease
1 Lambda HindIII
2 Lambda EcoRI& Hind III
3 Lambda EcoRI
4 Lambda BamHI
5 Lambda EcoRI & BamHI
6 Plasmid pUC18 EcoRI
7 Lambda Hae III
8 Calf Thymus EcoRI
9 Nucleosome  I  __________
10 Nucleosome II  __________

 

Plasmid DNA - pUC18

This DNA can be used in gene cloning experiments and will transform E. coli to an ampicillin resistant phenotype. This plasmid contains single cloning sites for a number of restriction endonucleases including EcoRI, BamHI, and Hind III.

3-14-30 30 µg (100 µl) $36.00
3-14-120 120 µg (400 µl) $62.61

Phage Lambda DNA

An excellent DNA preparation to show the specificity of restriction endonucleases

3-15-60  60 µg (150 µl) $25.64
3-15-240 240 µg (600 µl)  $65.01

Calf Thymus DNA

This DNA can be used to illustrate the G - C rich satellite sequences in the cow genome

3-16 5 mg (5 ml)  $36.00

Plasmid Restriction Fragment Set

An electrophoretic analysis of the four DNA samples provided with this set is shown on the agarose gel below. The ready-to-load DNA samples can be used as DNA size markers or in "DNA Fingerprinting" exercises designed by the instructor. Each of the four DNA samples contains sufficient material for 8 - 10 gel lanes and the fragments can readily be detected by methylene blue gel staining.

3-20  Restriction Fragment Set $69.05

Plasmid Restriction Fragment Set

 

 

 

 

 

 

 

 

Colony Transformation

a E. coli Transformation Kit

Complete ready-to-use kit for transformation of E. coli with plasmid pUC18. The success of the transformation is monitored by growing the bacteria on an ampicillin-containing medium. Sufficient materials supplied for sixteen platings. Kit includes E. coli, pUC18, CaCl2, nutrient broth, inoculating loops, pipets, tubes, petri dishes, and 400 ml of nutrient agar-ampicillin.

3-17 Transformation Kit $69.12

a Kits for DNA Isolation

Plasmid DNA Isolation Kit

Kit contains reagents needed to isolate plasmid DNA from E. coli. The procedure yields high-quality plasmid DNA that is suitable for restriction nuclease digestion. Sufficient materials are provided for about 30 "mini-preps" or 8 larger scale purifications. Kit includes lysis solution, SDS-NaOH solution, ammonium acetate, Tris-EDTA buffer, isopropanol, and an electrophoresis sample buffer.

3-18 Plasmid Isolation Kit $54.71

Genomic DNA Isolation Kit

The reagents in this kit are used to isolate DNA from bacteria or eukaryotic tissues. The unique procedure yields high quality genomic DNA without the use of toxic organic solvents. The procedure has been successfully used to isolate DNAs from E. coli, corn seedlings, wheat germ, chicken erythrocytes, and calf thymus. In each case, the DNAs were undergraded and were excellent substrates for a variety of restriction endonucleases. Kit includes NB-buffer, SDS, ammonium acetate, Tris-EDTA buffer, an electrophoresis buffer containing ribonuclease and wheat germ so that you can test the procedure. Ethyl alcohol and a centrifuge that can be operated at a force of at least 5000 x g are needed but not provided.

3-19 Genomic DNA Isolation Kit  $69.12

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Last modified: 10/02/07