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Second Strand cDNA Synthesis Kit
Second Strand cDNA Synthesis Kit
Invitrogen™

Second Strand cDNA Synthesis Kit

Invitrogen Second Strand cDNA Synthesis Kit is designed to produce double-stranded cDNA from the first-strand reaction without the need forRead more
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Catalog NumberNo. of ReactionsVector
A4857050 Reactions
A48571200 Reactions
Catalog number A48570
Price (USD)
1,190.65
Online Exclusive
1,264.00
Save 73.35 (6%)
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No. of Reactions:
50 Reactions
Price (USD)
1,190.65
Online Exclusive
1,264.00
Save 73.35 (6%)
Each
Add to cart
Invitrogen Second Strand cDNA Synthesis Kit is designed to produce double-stranded cDNA from the first-strand reaction without the need for intermediate organic extraction or ethanol precipitation steps. The convenient single-tube format speeds up the synthesis procedure and maximizes cDNA recovery. The kit contains premixed components to reduce the number of pipetting steps necessary to complete the procedure.

Features of the Second Strand cDNA Synthesis Kit include:
• Efficient synthesis of full-length double-stranded cDNA
• Fast—procedure completed in 1 hour
• Convenient—pre-mixed components
• Complete—includes residual RNA removal reagents

Applications
• Full-length double-stranded blunt-end cDNA synthesis for cloning
• Double-stranded cDNA library construction

WARNING: Cancer and Reproductive Harm – www.P65Warnings.ca.gov
Specifications
Final Product TypeDouble Stranded cDNA
Format50 reactions
No. of Reactions50 Reactions
Product TypecDNA Synthesis Kit
Quantity50 reaction
Reaction FormatSeparate Components
For Use With (Application)Cloning
Unit SizeEach
Contents & Storage
Contents:
• Second Strand Enzyme Mix, 1 mL
• 5X Second Strand Reaction Mix, 250 μL
• 0.5 M EDTA, pH 8.0, 1 mL
• RNase I, 10 U/μL, 500 μL
• Water, nuclease-free, 3 x 1.25 mL

Storage: -15 to -25°C

Frequently asked questions (FAQs)

What is the correct reaction temperature that I should use for second strand cDNA synthesis using the Second Strand cDNA Synthesis Kit?

The second strand reaction must be incubated at 16 degrees C to prevent spurious synthesis by DNA polymerase I. At higher temperatures, DNA polymerase I tends to strand-displace rather than nick translate.

When using the Second Strand cDNA Synthesis Kit, what steps do I need to take to properly prepare the second strand reaction mixture?

Dilute the first-strand reaction as outlined in the manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0019413_SecondStrandcDNASynthesisKit_UG.pdf) on page 2 under Second Strand cDNA Synthesis step. Failure to properly dilute the first-strand reaction changes the pH of the second strand reaction and influences the activity of the 3'-5' and 5'-3' exonuclease activities of DNA polymerase I.

When using the Second Strand cDNA Synthesis Kit, how can I avoid incomplete RT inactivation?

Always perform the RT inactivation step after first-strand cDNA synthesis.

I am getting low yield of second strand cDNA product using the Second Strand cDNA Synthesis Kit. What is the reason?

The main reasons are incomplete RT inactivation, improper preparation of the second strand reaction mixture, and incorrect reaction temperature.

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