DNA Staining Reagent

4,6-DiAmidino-2-PhenylIndole Dihydrochloride
C₁₆H₁₅N₅‧2(HCl)
M.W.: 350.25
Assay (HPLC): 98.8%
Solution: Clear (10g in 1L solution in water 20ºC)
TLC: Single Spot
Yellow Orange Powder

DAPI is an excellent dye for the staining of DNA. The most popular application of DAPI is its use as a reagent to detect mycoplasma or virus DNA in the cell culture.
Use 0.25mg/mL solution.

Stock Conc.: 1-2mg/mL, stable for 6-12 months when store at -20°C, room temp for 1-2 weeks, protect from light.
Working Conc.: 1:1000 dilution of stock solution

Russel, W.C. et al. (1975) Nature 253, 461-462
A simple cytochemical technique for demonstration of DNA in cells infected with mycoplasmas and viruses.

Store at 4ºC

DNA/RNA Experiment, For Nucleic Acid Hybridization

DNase, RNase not detected

Denhardt's solution is a blocking reagent for preventing the unspecific binding of nucleic acids to nitrocellulose or nylon membranes in hybridization experiments. Denatured DNA binds to nitrocellulose membranes, but not free RNA. Pretreatment of the membrane with Denhardt's solution prevents the binding of single stranded or (unspecific) denatured DNA.

The use of Denhardt's reagent is recommended for Northern hybridization, hybridization of RNA, 'single copy Southern hybridization' or hybridization of DNA, immobilized on nylon membranes.

Denhardt, D.T. (1966) Biochem. Biophys. Res. Com. 23, 641-646
A membrane-filter technique for the detection of complementary DNA.
Sambrook, J. et al, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory(1989), p 9.48-50, B15

Store at 4ºC

RNA Experiment, RNase Deactivator

DiEthyl PyroCarbonate 　
O(COOC₂H₅)₂
M.W.: 162.14
Assay: >99% Min.
Dimethylcarbonate: 1.0% Max.
Methanol: 0.2% Max.
Arsenic (As): 0.0002% Max.
Chloride (Cl): 0.001% Max.
Heavy Metal: 0.0005% Max.
Residue on Ignition: 0.1% Max.

Modification reagent for His and Tyr residues in proteins. In 0.1% solution, it inactivates RNase, thus protecting RNA from degradation.

For the treatment of water with DEPC, add 1 ml of DEPC to 1 liter bi-distilled water (0.1 % DEPC v/v) and stir over night. Autoclaving 20 minutes inactivates DEPC. DEPC reacts with water and hydrolyses to ethanol and CO₂.

Store at 4ºC

Caution: DEPC is suspected to be a carcinogen and should be handled with great care. Do not breathe fumes and avoid contact with skin! DEPC may damage eyes and mucous membranes!

Harmful, Irritant

Average Molecular Weight (Gel permeation chromatogryphy): 38,500 Dalton
Below 10,900 Dalton: 10%
Above 99,700 Dalton: 10%
pH: 6.1
Cloride (Cl): <100mg/kg
Nitrogen (N): <30mg/kg

Store at Room Temp.

Average Molecular Weight (Gel permeation chromatogryphy): 64,000 Dalton
Below 20,100 Dalton: 10%
Above 160,400 Dalton: 10%
pH: 6.4
Cloride (Cl): <100mg/kg
Nitrogen (N): <64mg/kg

Store at Room Temp.

DNA/RNA Experiment, For Nucleic Acid Hybridization

From dextran with an Av. M.W.: 500,000. Sodium salt, adjust to pH 7 with phosphate

Heparin-like polysaccharide, which improves the hybridization rate of nucleic acids.

Stock Conc.: 50%
Working Conc.: 10%

Store at 4ºC

DNA/RNA Experiment, DNA Size Marker
Datasheet

This ladder consists of forty blunt end DNA bands at exactly 100bp to 4,000bp in 100 bp increments. Two features of the ladder facilitate band identification:
1. A high intensity 500 bp band
2. Bands from 1,000bp to 4,000bp are more intense than bands from 100 bp to 900bp. G+C content of the bands is 48%. The DNA concentration of the stock solution is 1ug/uL. Thus, the average concentration of each band is about 28ng/uL.

3 Easy Steps: (For details, please refer to Datasheet)
1. Just add 50uL stock solution to 450uL ready-to -use loading dye.
2. Mix well.
3. Add 5-10uL per loading.

Store at -20ºC

DNA/RNA Experiment, DNA Size Marker
Datasheet

This ladder consists of 10 blunt ended DNA bands at exactly 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 kilobase. All bands (except 5 kb) are supplied at 30ng/uL, providing uniform staining intensity with ethidium bromide or sybr green I dyes. The 5kb band is supplied at triple the concentration, 90ng/uL, to provide an easy marker for band identification. All bands can be resolved using 0.6%-1.0% agarose gel electrophoresis.

3 Easy Steps: (For details, please refer to Datasheet)
1. Just add 50uL stock solution to 450uL ready-to -use loading dye.
2. Mix well.
3. Add 5-10uL per loading.

Store at -20ºC

DNA/RNA Experiment, DNA Size Marker

This ladder consists of 33 blunt ended DNA bands at exactly 200bp to 6600bp in 200bp increments. The high intensity 1000bp band facilitates band identification. G+C content of all band is 48%.

Store at -20ºC

DNA/RNA Experiment, DNA Size Marker

This ladder consists of 25 blunt ended DNA bands at exactly 20bp to 500bp in 20bp increments. The high intensity 100bp band facilitates band identification. G+C content of all bands is 50%.

Store at -20ºC

For nick translation, production of random fragments, cleavage of genomic DNA for footprinting, removal of DNA template after in vitro transcription, and removal of DNA from RNA samples prior to applications such as RT-PCR.

Heat Inactivation: 10 minutes at 65ºC in the presence of  Stop Solution.
Molecular Weight: 31,000 Daltons.
Source: Bovine pancreas.
This DNase solution does not contain an RNase inhibitor.

(RNA-Qualified) RNase-Free DNase is a DNase I (endonuclease) that degrades both double-stranded and single-stranded DNA, producing 3′-OH oligonucleotides. In the presence of Mg²⁺, DNase I attacks each strand of DNA independently, and the sites of cleavage are distributed in a statistically random fashion. In the presence of Mn²⁺, DNase I cleaves both strands of DNA at approximately the same site to yield fragments with blunt ends or protruding termini of one or two nucleotides in length.

Under different buffer conditions the amount of DNase required to completely digest a given amount of DNA may need to be empirically determined.

Store at –20ºC. Avoid exposure to frequent temperature changes.

Protein Experiment, Protein Disulfide Bond, Reducing Reagent

Dithioerythritol 　 　
C₄H₁₀O₂S₂
M.W.: 154.25
Assay: 99.2%
Oxidized DTE: 0.1g/100g
pH: 6.3

Dithioerythritol (DTE) is an isomer of dithiothreitol (DTT). In general, DTE and DTT are exchangeable.

Store at 4ºC

Protein Experiment, Protein Disulfide Bond, Reducing Reagent

Dithiothreitol
C₄H₁₀O₂S₂
M.W.: 154.25
99% Min
Melting Point: 41-42ºC
For Molecular Biology Research Use
Soln. (5% in water): Clear and Colorless

Dithiothreitol (DTT) is, like beta-mercaptoethanol, a reducing reagent for proteins and protects the cysteine residues against oxidation.

Stock Conc.: 1M in ddH₂O, Store at -20ºC
Working Conc.: 0.1-1mM

Cleland, W.W. (1964) Biochemistry 3, 480-482
Dithiothreitol, a new protective reagent for SH groups.

Store at 4ºC

Ready to use. Tested in SDS-PolyAcrylamide Gel Electrophoresis and Western blotting.

This Marker contains 10 proteins that resolve into sharp, tight bands. In Tris-Glycine 4-20% buffer, the bands are exactly 15, 20, 30, 40, 50, 60, 70, 90, 130, 175kDa. The 40 and 90kDa reference bands coupled with blue dye facilitate band identification. In Bis-Tris 10% MES buffer, the bands are exactly 14, 17, 25, 32, 43, 53, 62, 81, 125, 170kDa. The 32 and 81kDa reference bands coupled with blue dye facilitate band identification.

Recommendations for Loading:
1. Thaw the ladder either at room temperature or at 37-40ºC for a few minutes to dissolve precipitated solids. Do not boil.
2. Mix thoroughly, to ensure that the solution is homogeneous.
3. Load the following volumes of the ladder on SDS-polyacrylamide gel:
-10uL per well for mini gels, 5uL per well for blots.
-20uL per well for large gels, 10uL per well for blots.

Store at –20ºC for 12 months. Stable at 4ºC for 3 months.