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Anchored Oligo(dT)<sub>20</sub> Primer
Invitrogen™

Anchored Oligo(dT)20 Primer

Anchored Oligo(dT)20 Primer is a primer mixture consisting of a string of 20 deoxythymidylic acid residues followed by dV (dG, dA, or dC) and then by dN (dA, dT, dG, or dC).
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Catalog NumberQuantity
1257701120 μL
Catalog number 12577011
Price (USD)
215.65
Online exclusive
234.00
Save 18.35
Each
Quantity:
20 μL
Price (USD)
215.65
Online exclusive
234.00
Save 18.35
Each

Anchored Oligo(dT)20 Primer is a primer mixture consisting of a string of 20 deoxythymidylic acid residues followed by dV (dG, dA, or dC) and then by dN (dA, dT, dG, or dC). Due to the variable end-sequence, the primer is anchored to the 5'-end of the poly(A) tail of mRNA and prevents priming within the poly(A) tail.

Anchored Oligo(dT)20 Primer is recommended for use in cDNA labeling protocols and in RT-PCR. It is commonly used for first-strand cDNA systhesis.

Quality control: This product is qualified in a first-strand cDNA synthesis reaction

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

Specifications
5'Primer ModificationPhosphate
For Use With (Application)First-strand cDNA Synthesis
Primer Length22-mer
Primer Sequence5'd PO4 [(T)20VN]3'
Product TypePrimer
Purification MethodGel-purified, Desalted
Quantity20 μL
Shipping ConditionDry Ice
Concentration2.5 μg/μL
PrimerAnchored Oligo dT Primer
Unit SizeEach
Contents & Storage
• Anchored Oligo(dT)20 Primer in DEPC-treated water (20 μL at 2.5 μg/μL)

Store at –20°C.
Guaranteed stable for 6 months when properly stored.

Frequently asked questions (FAQs)

How does the Anchored Oligo(dT)20 Primer differ from standard oligo(dT) primers?

Anchored Oligo(dT) primers have 2 random bases at the 3' end of the stretch of Ts. The first random base is either an A, G, or C, while the second can be any of the 4 standard nucleotides, i.e., A, T, G or C. The use of anchored oligo(dT) primers results in increased cDNA synthesis yields.

For first-strand cDNA synthesis, is it better to use oligo(dT), random hexamers, gene-specific primer (GSP), or combination of these primers?

The choice of primer depends on your experimental goals. Oligo(dT) is recommended when using total RNA for cDNA synthesis. It is the key to full-length cDNA synthesis. Random hexamers give a series of short first-strand products spanning the entire mRNA. Use of random hexamers may be helpful if the PCR fragment is at the 5´ end of a large mRNA. To ensure full-length cDNA synthesis of large transcripts, oligo(dT) can be added along with random hexamers during first-strand synthesis. Gene-specific primers (GSP) for cDNA synthesis may also be used and are required in a few applications such as 5´ RACE and qRT-PCR. For GC-rich templates or templates rich in secondary structure, a GSP may not work as well as priming with oligo dT for first-strand synthesis. If an RT-PCR is problematic, trying different options of oligo dT, random primers and/or GSP for priming first-strand synthesis may resolve the issue. Oligo(dT)20 primer (Cat. No. 18418-020) is recommended for use with SuperScript III Reverse Transcriptase (Cat. no. 18080-044), ThermoScript Reverse Transcriptase (Cat. No. 12236-014), Thermo-X Reverse Trascriptase (Cat. No. 11150-025), and Cloned AMV Reverse Transcriptase (Cat. No. 12328-019).

Reference:
Frohman,M.A., Dush,M.K., Martin, G.R. (1988) Proc. Nat. Acad. Sci USA 85, 8998