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Invitrogen™ RNA 6000 Ladder

Catalog No. AM7152
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Catalog No. AM7152 Supplier Invitrogen™ Supplier No. AM7152
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Description

A set of six RNA transcripts with lengths of 0.2, 0.5, 1.0, 2.0, 4.0, and 6.0kb

The 0.2Kb band is at a concentration of 20ng/μL (A260). The RNA 6000 Ladder is designed for use with Agilent's lab-on-a-chip. Three tubes containing 20μL each are provided. 150ng (1μL) will generate 6 distinct bands when run on a RNA Labchip™ using the Total RNA or mRNA assay. Compatible with RNA LabChip.

DNA and RNA Purification and Analysis, Nucleic Acid Quantitation, RNA Quantitation

Order Info

Shipping Condition: Dry Ice

Specifications

Content And Storage Store at below –70°C.
Concentration 150 μg/ml
Ready to Load No
Size Range 0.2 to 6 kb
Gel Compatibility RNA LabChip
Quantity 3 tubes
Shipping Condition Dry Ice
Product Line Ambion
Product Type RNA 6000 Ladder

Frequently Asked Questions (FAQs)

Why are my RNA bands not sharp?

(1) RNA was not completely denatured. Electrophorese RNA under denaturing conditions. Use urea, formamide or formaldehyde gels, or glyoxal-treated RNA.

(2) For glyoxal-treated RNA, a buffer gradient formed during electrophoresis. Recirculate buffer during electrophoresis to prevent gradient formation.
What is the cause of extra bands when using an RNA ladder?

Extra bands appear in RNA Ladders for a few reasons:

(1) RNA was not completely denatured. Electrophorese RNA under denaturing conditions. Use urea, formamide or formaldehyde gels, or glyoxal-treated RNA.

(2) Extra bands may be a result of using formaldehyde that is not fresh; the pH becomes acidic in older formaldehyde. In our hands, when fresh formaldehyde with neutral pH was used, the extra bands disappeared.

(3) Alternatively, if the extra bands appear after hybridization, it could be that the gel purified probe contains contaminating vector DNA (pUC or pBR) that hybridizes to RNA Ladder template DNA.
Why are some of the RNA marker bands not visible?

Missing RNA bands may be due to:

(1) A small amount of RNA diffused out of gel during extended destaining. Minimize destaining time. Destaining for 2 hours is sufficient for most applications.

(2) RNA bands of similar molecular size were not resolved. Use the correct gel type and denaturing conditions.
Why are the RNA bands disappearing when looking and photographing a gel on a UV box?

RNA was exposed to UV light for extended periods of time. Minimize exposure to UV light. Stain and destain gels in the dark and photograph the gel immediately.
Why are the RNA marker bands so faint?

Many factors could affect the intensity of the bands as summarized below.

(1) Insufficient RNA was loaded on the gel. Increase the amount of RNA loaded.

(2) RNA was degraded. Avoid nuclease contamination of the RNA standards. Store RNA at -70 degrees C in formamide. Deionize formamide and glyoxal, store aliquots at -20 degrees C.

(3) RNA was electrophoresed off the gel. Electrophorese the gel for less time, at a lower voltage, or in a higher percentage gel.

(4) For ethidium bromide-stained RNA, improper UV light source was used. Use short-wavelength (254 nm) UV light. For ethidium bromide-stained RNA, improper staining and destaining conditions were used. Stain in the dark with 5 mg/mL ethidium bromide. For thin (3.2 mm) formaldehyde gels, stain 5 min and destain 1 h. For thicker gels, stain 30 min and destain 2 hr. To reduce background staining, use 0.66 M formaldehyde instead of 2.2 M formaldehyde in gels. For glyoxal RNA gels, stain and destain in 0.5 M ammonium acetate to help reduce background staining.

(5) For radiolabeled RNA, an improper labeling method was used.
What kind of RNA molecular weight ladders do you offer?

We offer the following RNA Ladders:


RiboRuler High Range RNA Ladder (Cat. No. SM1821)
RiboRuler Low Range RNA Ladder (Cat. No. SM1831)
Century Marker Templates, 100-500 bp (Cat. No. AM7780)
Century-Plus Marker Templates, 0.1-1 kb (Cat. No. AM7782)
Decade Markers, 10-150 bp (Cat. No. AM7778)
Millennium Markers-Formamide, 0.5-9 kb (Cat. No. AM7151)
RNA 6000 Ladder, 0.2-6 kb (Cat. No. AM7152)
RNA Century-Plus Markers, 0.1-1 kb (Cat. No. AM7145)
RNA Millennium Markers, 0.5-9 kb (Cat. No. AM7150)
What staining procedures are recommended for RNA ladders?

RNA can, in fact, be stained with ethidium bromide. However, proper staining procedures are important in order to visualize RNA. Insufficient staining and destaining will result in RNA bands being lost in the background. Ethidium bromide can be added to samples prior to electrophoresis.

We recommend the following amounts for this approach, using denaturing gels with formaldehyde contents of 2.2 M:
--For a gel thickness of 6.4 mm, the ethidium bromide should be added to the sample to a final concentration of 1.2 mg/mL.
--For a 3.2 mm thick gel, the final concentration should be 0.13 mg/mL.

If ethidium bromide staining is carried out after electrophoresis, the following recommendations should be followed (ethidium bromide at 5 mg/mL):
--3.2 mm thick gels containing 2.2 M formaldehyde, 5 min staining, 0.5 to 1 hr destaining
--6.4 mm thick gels containing 2.2 M formaldehyde, 5-30 min staining, 2 to 3 hr destaining
--6.4 mm thick gels containing 0.6 M formaldehyde, 10-20 min staining, 1 to 2 hr destaining

For Research Use Only. Not for use in diagnostic procedures.