Five Tips & Products for Avoiding Mishaps in Your Molecular Biology Applications

Tip 1: Agarose

Agarose gel electrophoresis remains the most widely used technique for separating nucleic acid fragments due to non-toxicity and a broad separation range. Following a few simple steps during gel casting can help you avoid burns and achieve best resolution.

  • Always use a flask 2–4 times the volume of the solution. Allow the powder to hydrate in the solution for a few minutes before heating. Weigh the flask and solution before and after heating and add warm distilled water to solution to bring it to its original weight.
  • To avoid bubble formation, cool the hot agarose solution to 50–60oC and pour carefully into gel cassette. Allow gel solution to cool gradually; rapid cooling will cause an irregular gel matrix and band distortion during electrophoresis.
  • When using low melting agarose gel, it is important to run the gel in a cold buffer. High voltage can cause overheating of the buffer, which can melt the gel. The UltraPure™ Agarose products are made from the highest purity biochemicals and packaged in a pouch that makes pouring and dispensing the powder easier—reducing the likelihood of spills and contamination.

Tip 2: Electrophoresis Buffer

The most commonly used buffers for DNA electrophoresis are TAE (Tris-acetate EDTA) or TBE (Tris-borate-EDTA). DNA fragments will migrate at different rates in these two buffers due to differences in ionic strength.

Typically, the TAE buffer provides better resolution of fragments greater than 4 kb, and the TBE buffer resolves 0.1–3 kb fragments. The TBE buffer is better suited for highvoltage (>150V) electrophoresis because of its higher buffering capacity and lower conductivity. If you mistakenly use water instead of buffer, there will essentially be no migration of DNA in the gel. Avoid mishaps with conveniently packaged, ready-to-use 10x TBE or 10x TAE Buffer.

Tip 3: Ultrapure Water

Laboratory water has multiple uses in the research labs, from glassware rinsing to highly sensitive applications involving biological samples. Unfortunately, in many labs, water contains many contaminants, such as trace organics, particles, bacteria, and, importantly, nucleases, all of which can have a detrimental effect on the resulting date.

For example, if nucleases are present in PCR reaction, they will effectively cleave any DNA, including the primers and probes. Amplification will therefore not occur. How pure is the source of water used in your PCR reaction? Why not eliminate possible contaminates in your PCR reaction by trying UltraPureTM DNase/RNase Free Distilled Water? With no DNase, RNase, or protease activity detected, it’s designed for use in all molecular biology applications.

Tip 4: DNA Ladders

An essential tool used during gel electrophoresis is a DNA marker or ladder, which is used to determine the size of an unknown DNA sample. DNA ladders are easy to identify because they contain regularly spaced, distinct-sized samples that, when run on an agarose gel, look like a ladder.

The recommended loading volume is dependent on detection methods: Typically, one ng DNA band can be detected with ethidium bromide staining. However, we normally load different volumes—e.g., 10 ul and 2 ul of a DNA ladder in the first and last lanes of a gel to orient the gel and to increase the quantity range. Use of two tracking dyes in the DNA ladders and samples ensures the DNA bands will not run off the gel and indicate maximum resolution has been achieved. Life Technologies offers a wide range of DNA ladders in the ready-to-load TrackItTM format. The TrackItTM DNA Ladders are ready to use—no need to heat, mix, or dilute.

Tip 5: DNA Stain

The most common dye used to make DNA or RNA bands visible is ethidium bromide (EtBr).

It fluoresces under UV light when the dye intercalates with DNA or RNA. Short exposure of nucleic acids to UV light can cause significant damage to the sample, which will reduce the efficiency of subsequent manipulations of samples, such as ligation or cloning.

If DNA is to be used after separation on agarose gel, it is best to avoid exposure to UV light by using a blue light excitation source and stain, such as SYBR Safe DNA Stain. SYBR Safe DNA Stain is a highly sensitive stain for visualization of DNA that has been shown to have a low level of mutagenicity and toxicity. It is classified as non-hazardous waste with similar sensitivity as EtBr.

For a limited time, we are offering UltraPure™ product essentials, DNA ladders and DNA stains at a special discounted price.

For UltraPure reagents, use promotion code MH0202 when placing your order*.
For DNA ladders, use promotion code P862959 when placing your order*.
For DNA stains, use promotion code P862950 when placing your order*.
Hurry! Offers expire September 14, 2012.

UltraPure™ reagents for electrophereisis | UltraPure™ reagents for protein isolation | UltraPure™ reagents for cloning | UltraPure™ reagents for hybridization | UltraPure™ reagents for general chemical | UltraPure™ reagents for nucleic acid purification | DNA ladders | DNA stains

UltraPure™ reagents for electrophereisis

SKU Product Catalog Size Discount  
15504-020 UltraPure™ Tris 1 kg Buy 1 get 1 free Add to cart
15505-035 UltraPure™ Urea 500 g Buy 1 get 1 free Add to cart
15505-050 UltraPure™ Urea 2 kg Buy 1 get 1 free Add to cart
15506-017 UltraPure™ Tris Hydrochloride 500 g Buy 1 get 1 free Add to cart
15512-023 UltraPure™ Acrylamide 500 g Buy 1 get 1 free Add to cart
15516-024 UltraPure™ N,N'-Methylenebisacrylamide 100 g Buy 1 get 1 free Add to cart
15525-017 UltraPure™ Sodium Dodecyl Sulfate (SDS) 500 g Buy 1 get 1 free Add to cart
15527-013 UltraPure™ Glycine 500 g Buy 1 get 1 free Add to cart
15553-027 UltraPure™ 10% SDS Solution 4 × 100 ml Buy 1 get 1 free Add to cart
15558-026 UltraPure™ 10X TAE Buffer 4 L Buy 1 get 1 free Add to cart
15558-042 UltraPure™ 10X TAE Buffer 1 L Buy 1 get 1 free Add to cart
15567-027 UltraPure™ 1M Tris-HCI, pH 7.5 1 L Buy 1 get 1 free Add to cart
15568-025 UltraPure™ 1M Tris-HCI, pH 8.0 1 L Buy 1 get 1 free Add to cart
15575-020 UltraPure™ 0.5M EDTA, pH 8.0 4 × 100 ml Buy 1 get 1 free Add to cart
15576-028 UltraPure™ Ethylenediaminetetraacetic Acid, Disodium Salt, Dihydrate (Na2EDTA•2H2O) 500 g Buy 1 get 1 free Add to cart
15581-028 UltraPure™ 10X TBE Buffer 10 L Buy 1 get 1 free Add to cart
15581-044 UltraPure™ 10X TBE Buffer 1 L Buy 1 get 1 free Add to cart
15585-011 UltraPure™ 10 mg⁄ml Ethidium Bromide 10 ml Buy 1 get 1 free Add to cart
16500100 UltraPure™ Agarose 100 g Buy 1 get 1 free Add to cart
16500500 UltraPure™ Agarose 500 g Buy 1 get 1 free Add to cart
16520050 UltraPure™ Low Melting Point Agarose 50 g Buy 1 get 1 free Add to cart
16520100 UltraPure™ Low Melting Point Agarose 100 g Buy 1 get 1 free Add to cart
16550100 UltraPure™ Agarose-1000 100 g Buy 1 get 1 free Add to cart
24710-030 UltraPure™ DNA Typing Grade® 50X TAE Buffer 1 L Buy 1 get 1 free Add to cart
24730-020 UltraPure™ 10% SDS Solution 1 L Buy 1 get 1 free Add to cart
24740-011 UltraPure™ 5M NaCl 10 L Buy 1 get 1 free Add to cart
LC6675 Novex® TBE Running Buffer (5X) 1 L Buy 1 get 1 free Add to cart
LC6876 Novex® TBE-Urea Sample Buffer (2X) 10 ml Buy 1 get 1 free Add to cart
R-33400 Rhinohide™ Polyacrylamide Gel Strengthener Concentrate 200 ml Buy 1 get 1 free Add to cart

UltraPure™ reagents for protein isolation

SKU Product Catalog Size Discount  
15503-022 UltraPure™ Sucrose 5 kg Buy 1 get 1 free Add to cart
15508-013 UltraPure™ Dithiothreitol 5 g Buy 1 get 1 free Add to cart
15524-010 UltraPure™ TEMED 30 ml Buy 1 get 1 free Add to cart
15525-017 UltraPure™ Sodium Dodecyl Sulfate (SDS) 500 g Buy 1 get 1 free Add to cart
15527-013 UltraPure™ Glycine 500 g Buy 1 get 1 free Add to cart
15553-027 UltraPure™ 10% SDS Solution 4 × 100 ml Buy 1 get 1 free Add to cart
24730-020 UltraPure™ 10% SDS Solution 1 L Buy 1 get 1 free Add to cart
R-33400 Rhinohide™ Polyacrylamide Gel Strengthener Concentrate 200 ml Buy 1 get 1 free Add to cart

UltraPure™ reagents for cloning

SKU Product Catalog Size Discount  
10977-015 UltraPure™ DNase/RNase-Free Distilled Water 500 ml Buy 1 get 1 free Add to cart
10977-023 UltraPure™ DNase/RNase-Free Distilled Water 10 × 500 ml Buy 1 get 1 free Add to cart
15504-020 UltraPure™ Tris 1 kg Buy 1 get 1 free Add to cart
15506-017 UltraPure™ Tris Hydrochloride 500 g Buy 1 get 1 free Add to cart
15567-027 UltraPure™ 1M Tris-HCI, pH 7.5 1 L Buy 1 get 1 free Add to cart
15568-025 UltraPure™ 1M Tris-HCI, pH 8.0 1 L Buy 1 get 1 free Add to cart
16500100 UltraPure™ Agarose 100 g Buy 1 get 1 free Add to cart
16500500 UltraPure™ Agarose 500 g Buy 1 get 1 free Add to cart
16520050 UltraPure™ Low Melting Point Agarose 50 g Buy 1 get 1 free Add to cart
16520100 UltraPure™ Low Melting Point Agarose 100 g Buy 1 get 1 free Add to cart
16550100 UltraPure™ Agarose-1000 100 g Buy 1 get 1 free Add to cart

UltraPure™ reagents for hybridization

SKU Product Catalog Size Discount  
15525-017 UltraPure™ Sodium Dodecyl Sulfate (SDS) 500 g Buy 1 get 1 free Add to cart
15553-027 UltraPure™ 10% SDS Solution 4 × 100 ml Buy 1 get 1 free Add to cart
15557-036 UltraPure™ 20X SSC 4 L Buy 1 get 1 free Add to cart
15557-044 UltraPure™ 20X SSC 1 L Buy 1 get 1 free Add to cart
15591-027 UltraPure™ 20X SSPE 10 L Buy 1 get 1 free Add to cart
15591-043 UltraPure™ 20X SSPE 1 L Buy 1 get 1 free Add to cart
15632-011 UltraPure™ Salmon Sperm DNA Solution 5 × 1 ml Buy 1 get 1 free Add to cart
15633-019 UltraPure™ Calf Thymus DNA Solution 5 × 1 ml Buy 1 get 1 free Add to cart
15634-017 UltraPure™ Herring Sperm DNA Solution 5 × 1 ml Buy 1 get 1 free Add to cart
24730-020 UltraPure™ 10% SDS Solution 1 L Buy 1 get 1 free Add to cart
750018 50X Denhardt Solution 100 ml Buy 1 get 1 free Add to cart

UltraPure™ reagents for general chemical

SKU Product Catalog Size Discount  
15507-023 UltraPure™ Cesium Chloride, Optical Grade 1 kg Buy 1 get 1 free Add to cart
15514-011 UltraPure™ Glycerol 500 ml Buy 1 get 1 free Add to cart
15514-029 UltraPure™ Glycerol 1 gal Buy 1 get 1 free Add to cart
15515-026 UltraPure™ Formamide 500 g Buy 1 get 1 free Add to cart

UltraPure™ reagents for nucleic acid purification

SKU Product Catalog Size Discount  
10813-012 UltraPure™ DEPC-treated Water 4 × 1.25 ml Buy 1 get 1 free Add to cart
10814-010 UltraPure™ Glycogen 100- µl Buy 1 get 1 free Add to cart
10977-015 UltraPure™ DNase/RNase-Free Distilled Water 500 ml Buy 1 get 1 free Add to cart
10977-023 UltraPure™ DNase/RNase-Free Distilled Water 10 × 500 ml Buy 1 get 1 free Add to cart
15502-016 UltraPure™ Guanidine Hydrochloride 500 g Buy 1 get 1 free Add to cart
15509-037 UltraPure™ Phenol 500 g Buy 1 get 1 free Add to cart
15513-039 UltraPure™ Buffer-Saturated Phenol 100 ml Buy 1 get 1 free Add to cart
15513-047 UltraPure™ Buffer-Saturated Phenol 400 ml Buy 1 get 1 free Add to cart
15514-011 UltraPure™ Glycerol 500 ml Buy 1 get 1 free Add to cart
15514-029 UltraPure™ Glycerol 1 gal Buy 1 get 1 free Add to cart
15577-018 UltraPure™ Guanidine Isothiocyanate Solution 500 ml Buy 1 get 1 free Add to cart
15593-031 UltraPure™ Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) 100 ml Buy 1 get 1 free Add to cart
15593-049 UltraPure™ Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) 400 ml Buy 1 get 1 free Add to cart
15594-047 UltraPure™ Phenol:Water (3.75:1, v/v) 400 ml Buy 1 get 1 free Add to cart
16520050 UltraPure™ Low Melting Point Agarose 50 g Buy 1 get 1 free Add to cart
16520100 UltraPure™ Low Melting Point Agarose 100 g Buy 1 get 1 free Add to cart
750023 UltraPure™ DEPC-Treated Water 1 L Buy 1 get 1 free Add to cart
750024 UltraPure™ DEPC-Treated Water 4 × 100 ml Buy 1 get 1 free Add to cart
LC6675 Novex® TBE Running Buffer (5X) 1 L Buy 1 get 1 free Add to cart
LC6876 Novex® TBE-Urea Sample Buffer (2X) 10 ml Buy 1 get 1 free Add to cart

DNA ladders

SKU Product Catalog Size Discount  
10068-013 Low DNA Mass™ Ladder 50 applications 25% Add to cart
10416-014 50 bp DNA Ladder 50 µg 25% Add to cart
10488-019 TrackIt™ 10 bp DNA Ladder 20 applications 25% Add to cart
10488-022 TrackIt™ 25 bp DNA Ladder 20 applications 25% Add to cart
10488-037 TrackIt™ X174RF DNA/Hae III Fragments 100 applications 25% Add to cart
10488-043 TrackIt™ 50 bp DNA Ladder 100 applications 25% Add to cart
10488-058 TrackIt™ 100 bp DNA Ladder 100 applications 25% Add to cart
10488-064 TrackIt™ λ DNA/Hind III Fragments 100 applications 25% Add to cart
10488-085 TrackIt™ 1 Kb Plus DNA Ladder 100 applications 25% Add to cart
10496-016 High DNA Mass™ Ladder 50 applications 25% Add to cart
10511-012 1 Kb DNA Extension Ladder 100 µg 25% Add to cart
10596-013 250 bp DNA Ladder 75 µg 25% Add to cart
10597-011 25 bp DNA Ladder 50 µg 25% Add to cart
10787-018 1 Kb Plus DNA Ladder 250 µg 25% Add to cart
10787-026 1 Kb Plus DNA Ladder 1000 µg 25% Add to cart
10821-015 10 bp DNA Ladder 50 µg 25% Add to cart
15611-015 X174 RF DNA/Hae III Fragments 40 µg 25% Add to cart
15612-013 λ DNA/Hind III Fragments 500 µg 25% Add to cart
15613-011 123 bp DNA Ladder 100 µg 25% Add to cart
15613-029 123 bp DNA Ladder 250 µg 25% Add to cart
15622-012 Supercoiled DNA Ladder 25 µg 25% Add to cart
15628-019 100 bp DNA Ladder 50 µg 25% Add to cart
15628-050 100 bp DNA Ladder 250 µg 25% Add to cart

DNA stains

SKU Product Catalog Size Discount  
15585-011 UltraPure™ 10 mg⁄ml Ethidium Bromide 10 ml 20% off Add to cart
S-11494 SYBR® Gold Nucleic Acid Gel Stain (10,000X Concentrate in DMSO) 500- µl 15% off Add to cart
S33100 SYBR® Safe DNA Gel Stain in 0.5X TBE, 1 L 1 L 15% off Add to cart
S33101 SYBR® Safe DNA Gel Stain in 0.5X TBE, 4 L 4 L 15% off Add to cart
S33102 SYBR® Safe DNA Gel Stain 400- µl 15% off Add to cart
S33110 SYBR® Safe DNA Gel Stain Starter Kit 1 个 15% off Add to cart
S33111 SYBR® Safe DNA Gel Stain in 1X TAE, 1 L 1 L 15% off Add to cart
S33112 SYBR® Safe DNA Gel Stain in 1X TAE, 4 L 4 L 15% off Add to cart
S-7563 SYBR® Green I Nucleic Acid Gel Stain - 10,000X concentrate in DMSO, 500 µL 500- µl 15% off Add to cart
S-7567 SYBR® Green I Nucleic Acid Gel Stain, 10,000X concentrate in DMSO (1 ml) 1- ml 15% off Add to cart
S-7585 SYBR® Green I Nucleic Acid Gel Stain, 10,000X concentrate in DMSO - Special Packaging 20 × 50 µl 15% off Add to cart

 

Terms and Conditions:

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.

*This promotion is open to academic end users in Hong Kong from July 13 to September 14, 2012. Discount will apply to orders received by Life Technologies Limited no later than September 14, 2012 or until promotional supplies are depleted, whichever comes first. Discount applies to list price in effect at the time order is received by Life Technologies. Cannot be combined with other discounts or promotions. Offer void where prohibited, licensed or restricted by federal, state, provincial, or local laws or regulation or agency/institutional policy. Other restrictions may apply. Life Technologies Limited reserves the rights at its sole discretion to determine the eligibility to receive the special offer.