Search

  • お問い合わせ一覧​
  • クイックオーダー

E-Gel™ CloneWell™ Agarose Gels with SYBR Safe™ DNA Gel Stain, 0.8% + E-Gel™ iBase™ Power System + E-Gel™ Safe Imager™ Transilluminator Starter Kit - FAQs

View additional product information for E-Gel™ CloneWell™ Agarose Gels with SYBR Safe™ DNA Gel Stain, 0.8% + E-Gel™ iBase™ Power System + E-Gel™ Safe Imager™ Transilluminator Starter Kit - FAQs (G6500ST)

11 product FAQs found

TaqMan Assayでジェノタイピングを行う場合、MGBプローブとTAMRAプローブのどちらが適しているでしょうか。

どちらのプローブもご利用いただけます。ただ、ジェノタイピングアッセイを行う場合にはMGBプローブの方が いくつかメリットがございます。 MGBプローブはMinor groove-binderを3’末端に持っており、この効果でプローブのTm値が上がります。 そのため、全く同じ配列でMGBプローブとTAMRAプローブを合成・比較した場合、MGBプローブの方が Tm値が高くなります。つまり、至適TmにするのにMGBプローブの方が短い配列で済むということです。 この差がジェノタイピングアッセイのメリットになります。 プローブの配列が短かくなるほど、完全マッチとミスマッチの配列でTm値の変動が大きくなります。 この効果でミスマッチ配列をより検出しにくくなるため、SNPのような1塩基の違いも特異性高く識別 できます。 また、MGBプローブは無蛍光のクエンチャーを使用しているため、バックグランド蛍光が低く抑えられています。 この効果もアレル識別に貢献しています。

TaqMan® Gene Expression Assaysのチューブラベルには何の日付が書いてありますか。

使用期限が記載されています。適切な保存環境で5年ですが、保証期間は未開封の状態で1年です。

The E-Gel Safe Imager Real-Time Transilluminator for the iBase Power System is not working. What should I do?

Please check the cord you are using for the iBase Power System. Oftentimes, the power cords for the PowerBase v.4 system and the E-Gel Safe Imager Real-Time Transilluminator get mixed up. If this is the case, the blue light will not come on even though the fan and LED light will operate. You will need to use the iBase cord, which should say 48 V on it.

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

How can I get better separation of my bands?

First check the percentage of your agarose gel. A higher percentage will help you to resolve smaller molecular weights while a lower percentage will help you to resolve larger molecular weights.

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

What is the difference between the E-Gel Safe Imager Real-Time Transilluminator (Cat. No. G6500) and the Safe Imager 2.0 Blue-Light Transilluminator (Cat. No. G6600)?

The E-Gel Safe Imager Real-Time Transilluminator docks onto the E-Gel iBase Power System and allows real-time imaging of the migration of DNA/RNA in E-Gel Agarose Gels containing SYBR Safe stain. The Safe Imager 2.0 Blue-Light Transilluminator is an independent system designed for viewing stained gels on the benchtop.

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

How much DNA can I load for my E-Gel Agarose Gels?

To determine the amount of DNA to load per well for your specific E-Gel agarose gel, please refer to the table on page 14 of the E-Gel Power Snap Electrophoresis System user guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017050_egel_powersnapsystem_UG.pdf).

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

Is it important to include water in the second row of wells for the E-Gel SizeSelect Agarose Gels and E-Gel CloneWell Agarose Gels?

Yes, please ensure that the second row is filled with sterile water prior to running your band of interest into the collection well. Please note that the refill volume may vary between wells. Do not overfill.

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

I would like to isolate DNA after gel electrophoresis. What do you recommend?

We offer our E-Gel SizeSelect Agarose Gels as well as our E-Gel CloneWell Agarose Gels, which are double-comb, precast agarose gels with simplified DNA recovery. Load your sample into the top row and electorphorese until your band or desired size range enters the bottom row. Then, easily remove the size-selected DNA with a pipette. No additional gel purification steps are necessary.

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

What does intact RNA look like when run on an agarose gel?

Intact RNA should have a 2:1 ratio of 28S:18S bands. You may see a smear of RNA that extends from <9 kb to 0.5 kb, indicating the presence of mRNA in the sample. To see an image or to read more about RNA assessment, visit this website (https://www.thermofisher.com/us/en/home/references/protocols/nucleic-acid-purification-and-analysis/rna-protocol/agarose-gel-electrophoresis-of-rna.html).

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

How can I perform RNA electrophoresis?

For nondenaturing RNA electrophoresis, we recommend using our E-Gel Precast Agarose Gels. Please note that E-Gel Agarose Gels are not validated to be RNAse-free. However, many of our customers routinely use E-Gel Agarose Gels for RNA analysis with success. If RNA is run on an E-Gel Agarose Gel, any loading buffer that would be used for nondenaturing RNA electrophoresis should be fine.

For denaturing RNA electrophoresis, there are several denaturing agents to choose from, including formaldehyde, glyoxal, formamide, and methyl mercury. Denaturing conditions disrupt hydrogen bonding so that RNA runs without secondary structure, as single-stranded molecules.

For denaturing RNA electrophoresis, our E-Gel EX Agarose Gels can be used. The only denaturing agent that is compatible with the E-Gel EX system is formamide, 50-90%. Using other denaturing agents will result in poor band separation and morphology. Please note that we do not recommend running samples prepared in RNA loading buffer on the same gel with samples prepared in water. Please see below for the RNA loading buffer recipe and denaturing electrophoresis conditions:

RNA Loading Buffer:
Deionized formamide: 200 µL
10X MOPS-EDTA-Sodium Acetate Buffer (0.4 M MOPS, pH 7.0, 0.1 M sodium acetate, 10 mM EDTA): 40 µL
Deionized formaldehyde: 76 µL
Water: 14 µL

Denaturing Electrophoresis Conditions:
1. Mix 15 µL of RNA loading buffer with 1-5 µL of RNA (1-5 µg).
2. Heat samples at 65 degrees C for 10 min to denature RNA.
3. Place samples on ice immediately after heating.
4. Load entire sample onto an E-Gel EX agarose gel.
5. Electrophorese for 30 minutes.

For denaturing RNA electrophoresis under formaldehyde-free conditions, we recommend using our NorthernMax-Gly Kit (Cat. No. AM1946). With this kit, RNA samples are denatured in glyoxal/DMSO loading buffer and run on a glyoxal-containing agarose gel.

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

Should I use agarose or polyacrylamide gels for DNA electrophoresis?

Agarose is commonly used as it is nontoxic, easy to use, and offers a broad range of separation. We offer precast E-Gel Agarose Gels or reagents to pour your own agarose gels. Polyacrylamide gels are typically used for high resolution of DNA molecules that range in size from 10-3,000 bp. We offer precast Invitrogen TBE polyacrylamide gels and UltraPure reagents.

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.