Call us toll-free

Quick academic help

Don't let the stress of school get you down! Have your essay written by a professional writer before the deadline arrives.

Calculate the price

Pages:

275 Words

$19,50

Evrogen Mint-2 cDNA synthesis kit: Method overview

Hi all,

May I know if any of the forumers used kits to synthesize ds cDNA before. If yes, which kits or what type of kits that you are using? I am trying to look for one, that can synthesize around 4-5kb length fRNA. Had one 1st strand cDNA synthesis kit, but it's not suitable for me because I need a ds cDNA to do RE digestion and cloning.

Needed advice and suggestion/opinion regarding to kits or methods or protocol.

Thanks in advance.

Cheerioet.

ds cDNA synthesis kit - PCR Reagents and Equipments
Photo provided by Pexels

SMART (Switching Mechanism at 5’ End of RNA Template) is a unique technology that allows the efficient incorporation of known sequences at both ends of cDNA during first strand synthesis, without adaptor ligation. The presence of these known sequences is crucial for a number of downstream applications including amplification, RACE, and library construction. While a wide variety of technologies can be employed to take advantage of these known sequences, the simplicity and efficiency of the single-step SMART process permits unparalleled sensitivity and ensures that full-length cDNA is generated and amplified.

cDNA Synthesis Kits - SMART cDNA Synthesis - Clontech

cDNA prepared with Mint-2 kit can be normalized using  (Cat# NK003) to decrease the prevalence of high abundant transcripts.
Photo provided by Flickr

One of the greatest advantages of SMART technology is its increased efficiency compared to traditional technologies such as adaptor ligation. Its high efficiency and sensitivity enables you to use a very limited quantity of starting material, such as microdissected tissues, laser-captured cells, biopsy samples, etc. As little as 1-2 ng of total RNA is sufficient for generating a highly representative cDNA pool for different downstream applications.

Because cDNA synthesis is susceptible to interruption by secondary structures in the template RNA, the 5' ends of genes are typically underrepresented in cDNAs synthesized by conventional methods. During SMART cDNA synthesis truncated products resulting from premature termination of the reverse transcription reaction do not incorporate the SMART(er) oligonucleotide and consequently are not amplified during PCR. Thus, cDNA created using our SMART technology and amplified by long-distance PCR is enriched for full-length cDNA. Furthermore, because the 5’ SMART(er) sequence and modified oligo dT primer are not added onto genomic DNA or cDNA transcribed from ribosomal RNA, cDNA that is generated using SMART is free of these contaminating agents.

PrimeScript ds cDNA Synthesis Kit ; ..

Hi all, May I know if any of the forumers used kits to synthesize ds cDNA before
Photo provided by Pexels

cDNA synthesis method utilizes a specific feature of the Moloney murine leukemia virus reverse transcriptase (RT). First-strand cDNA synthesis starts from the 3'-end CDS adapter containing an oligo(dT) sequence that anneals to poly(A) stretches of RNA. When RT reaches the 5'-end of the mRNA, it adds several non-template nucleotides, primarily deoxycytidines, to the 3'-end of the newly synthesized first-strand cDNA (Schmidt and Mueller, 1999; Matz , 1999). This oligo(dC) stretch base anneals to a complementary oligo(dG) sequence located at the 3'-end of a special deoxyribooligonucleotide adapter, called PlugOligo. Mint RT identifies PlugOligo as an extra part of the RNA template and continues first strand cDNA synthesis to the end of the oligonucleotide, thus incorporating PlugOligo sequence into the 5'-end of cDNA.

Mint-2 cDNA synthesis kit is designed to synthesize full-length-enriched double stranded (ds) cDNA from total or poly(A)+ RNA. Synthesized cDNA can be used for construction of cDNA libraries, subtractive hybridization (SSH), high-throughput sequencing on the next generation sequencing platforms (Roche/454, ABI/SOLiD or Illumina/Solexa), and other applications.

Evrogen Mint-2 cDNA synthesis kit: Summary
Photo provided by Flickr
Order now
  • UNMATCHED QUALITY

    As soon as we have completed your work, it will be proofread and given a thorough scan for plagiarism.

  • STRICT PRIVACY

    Our clients' personal information is kept confidential, so rest assured that no one will find out about our cooperation.

  • COMPLETE ORIGINALITY

    We write everything from scratch. You'll be sure to receive a plagiarism-free paper every time you place an order.

  • ON-TIME DELIVERY

    We will complete your paper on time, giving you total peace of mind with every assignment you entrust us with.

  • FREE CORRECTIONS

    Want something changed in your paper? Request as many revisions as you want until you're completely satisfied with the outcome.

  • 24/7 SUPPORT

    We're always here to help you solve any possible issue. Feel free to give us a call or write a message in chat.

Order now

If yes, which kits or what type of kits that you are using

At the final step, ds cDNA is amplified by PCR. Use of Encyclo polymerase and specially designed primers allows synthesis of full-length-enriched cDNA that is flanked by PlugOligo and CDS adapter sequences.

TeloPrime Full-Length cDNA Amplification Kit | Lexogen

Mint-2 cDNA synthesis kit is designed to synthesize full-length-enriched double stranded (ds) cDNA from total or poly(A)+ RNA. Synthesized cDNA can be used for construction of cDNA libraries, subtractive hybridization (SSH), high-throughput sequencing on the next generation sequencing platforms (Roche/454, ABI/SOLiD or Illumina/Solexa), and other applications.

The TeloPrime Full-Length cDNA Amplification Kit is a very 5 ..

You may not need a kit to do second strand cDNA synthesis. After first strand cDNA synthesis, you just add DNA polymerase I, RNAase H (see this )



full-length cDNA synthesis is ..

During first-strand SMARTer cDNA synthesis known universal primer sequences are incorporated at both ends of the cDNA. Following first-strand synthesis, SMARTer cDNA is immediately available for PCR amplification.

Order now
  • You submit your order instructions

  • We assign an appropriate expert

  • The expert takes care of your task

  • We send it to you upon completion

Order now
  • 37 684

    Delivered orders

  • 763

    Professional writers

  • 311

    Writers online

  • 4.8/5

    Average quality score

Order now
  • Kim

    "I have always been impressed by the quick turnaround and your thoroughness. Easily the most professional essay writing service on the web."

  • Paul

    "Your assistance and the first class service is much appreciated. My essay reads so well and without your help I'm sure I would have been marked down again on grammar and syntax."

  • Ellen

    "Thanks again for your excellent work with my assignments. No doubts you're true experts at what you do and very approachable."

  • Joyce

    "Very professional, cheap and friendly service. Thanks for writing two important essays for me, I wouldn't have written it myself because of the tight deadline."

  • Albert

    "Thanks for your cautious eye, attention to detail and overall superb service. Thanks to you, now I am confident that I can submit my term paper on time."

  • Mary

    "Thank you for the GREAT work you have done. Just wanted to tell that I'm very happy with my essay and will get back with more assignments soon."

Ready to tackle your homework?

Place an order