It is important to use highquality rna for your cdna prepar ation. I want to synthesize cdna for qpcr using the iscript cdna synthesis kit. Mar 11, 2012 rna isolation from embryonic zebrafish and cdna synthesis for gene expression analysis samuel m. The first strand cdna synthesis kit is used for the synthesis of the first strand cdna as the starting reaction for rtpcr. Readyscript cdna synthesis mix complete reagent for first. This kit includes just three tubes comprehensive of the reagents. However, this step can be omitted in some cases unpublished results. The kit is formatted for highyield production of cdna up to 12 kb and each reaction accommodates a range of rna amounts from fg to g. This robust reverse transcription kit allows a selection of.
Storage and stability store the iscript cdna synthesis kit at 20 c in a constant temperature freezer. At just minutes, the lunascript rt supermix kit offers the shortest available firststrand cdna synthesis protocol comparison of recommended protocols for cdna synthesis. The kit utilizes our smart switching mechanism at 5 end of rna template technology coupled with pcr amplification to generate high yields of fulllength doublestranded cdna suitable for various applications, including clontech pcrselect cdna subtraction, nondirectional cloning, and preparation of. The premium quality reverse transcriptase, ultrapure. This 5x concentrated mix provides all necessary components except rna template for first. Affinityscript qpcr cdna synthesis kit provides a streamlined master mix format and fast protocol, with a 15minute cdna synthesis step and an overall protocol time of just 25 minutes. Firststrand synthesis is performed in triplicate on each dilution of a logfold serial dilution of hela cell total rna from 1 pg to 1 g. The kit is supplied in a 2tube format with the vilo reaction mix and an enzyme blend in. Freeman school of health sciences, purdue university. Random random primers ensure that the first strand synthesis. Accuscript high fidelity 1st strand cdna synthesis kit. Mmulv enzyme mix combines mmulv reverse transcriptase and murine.
Superscript vilo cdna synthesis kit product information sheet. The monsterscript 1ststrand cdna synthesis kit has been discontinued. The iscript cdna synthesis kit from biorad provides a simple means of producing cdna for cloning purposes or realtime pcr. This kit is intended for use by researchers using biorads iscript cdna synthesis kit previously developed and manufactured by quanta for ongoing studies or established standard operating. Standard operating procedure ucsf helen diller family. The smarter pcr cdna synthesis kit is an improved version of. Random random primers ensure that the first strand synthesis occurs efficiently with all species of rna molecules present, including mrna and rrna. Onetenth of each firststrand reaction is used for qpcr amplification. The novel qscript reaction mix provides all the necessary components for cdna synthesis except enzyme and rna template. It can be used with either sequencespecific primers, polydt15 primers, or. He also mentioned that when they hear them disputes over difering on enoch, the. The optimized 5x transamp buffer includes all necessary components for cdna synthesis, including a unique blend of oligo dt and random hexamer primers and the ribosafe rnase inhibitor.
This 5x concentrated mix provides all necessary components except rna template for firststrand synthesis including. Read the safety data sheets sdss and follow the handling. The enzymes lack of rnase h activity contributes to its ability to make longer cdnas and more complete fulllength libraries of firststrand cdna. Highcapacity cdna reverse transcription kits product. The premium quality reverse transcriptase, ultrapure dntps and an optimized reaction buffer ensure superior results with highest reproducibility. Some couple cdna synthesis and quantitative realtime pcr in a single reaction. Recombinant mmulv reverse transcriptase generates cdna up to 10 kb supplied with oligodt primers, randomized primer mix, and nucleasefree water. Enter synonyms, trade names or additional information found in section 1. The kit is supplied in a 2tube format with the vilo reaction mix and an enzyme blend in separate tubes. Truscript first strand cdna synthesis kit product insert. It can be used with either sequencespecific primers, polydt15 primers, or random primers, pdn6.
This twotube kit is optimized to yield sensitive, unbiased representation over a broad dynamic range, with minimal setup and reaction time. Protocol for cdna synthesis and qrtpcr cdna synthesis. Highquality rna can be obtained by isolating total rna. The lunascript rt supermix kit requires the shortest reaction time and tolerates elevated temperatures, reducing complications from rna secondary structure. We recommend incubation at 42c for one hour for maximum yield and length. This twotube kit is optimized to yield sensitive, unbiased. The qscript cdna synthesis kit is a sensitive and easytouse solution for rna quantification using twostep rtpcr. The protoscript first strand cdna synthesis kit is contains everything needed for cdna synthesis. Tetro cdna synthesis kit contains our highly sensitive mmlv reverse transcriptase, oligo dt 18 and random hexamer primers and all the necessary components to. Storage and stability store the iscript cdna synthesis kit at 20c in a constanttemperature freezer. This kit is intended for use by researchers using biorads iscript cdna synthesis kit previously developed and manufactured by quanta for ongoing studies or established stan dard operating procedures that cannot be readily switch to qscript cdna supermix.
General description readyscript cdna synthesis mix is a sensitive and easytouse solution for twostep rtpcr. When stored under these conditions, the kit components are stable for a minimum of one year after ship date. The kit uses mmlv reverse transcriptase with lower rnase. The genome core has been using the biorad iscript cdna synthesis kit for at least two years, and has found it to be the best method in terms of ease, reliability. Hi, in our lab we study gene expression in plants and good rna and cdna synthesis is very important. It uses a blend of oligo dt and random hexamer primers, thus working well with a wide variety of targets. The qscript cdna synthesis kit is functionally tested in reverse transcription quantitative pcr rtqpcr. The superscript iii cellsdirect cdna synthesis kit is optimized for synthesizing firststrand cdna directly from a mammalian cell lysate without first isolating the rna. The novel qscript reaction mix provides all the necessary components for cdna. This twotube kit can accommodate dilute rna samples. This kit includes just three tubes comprehensive of reagents. Superscript vilo cdna synthesis kit product information. The tetro cdna synthesis kit is the more flexible solution. Rna isolation from embryonic zebrafish and cdna synthesis for gene expression analysis samuel m.
This gives the same relative representation in cdna templates, regardless of gene abundance, making it excellent for use in qpcr studies. Efficiency kits varied in their ability to efficiently generate. The iscript cdna synthesis kit is a sensitive and easytouse firststrand cdna synthesis kit for gene expression analysis using realtime qpcr. Lysis and reverse transcription are performed in the same tube, and the resulting firststrand cdna is ready to use in cloning and pcr. The user can use their own gene specific primers or either the randomers or oligodt included separately in the kit. The kit utilizes our smart switching mechanism at 5 end of rna template technology coupled with pcr. This is recommended when the cdna synthesis reaction uses less than 10 ng of rna input.
Script cdna synthesis kit, reverse transcriptases with. You selects the desired setup, especially the desired rt primer. The sensifast cdna synthesis kit displays excellent linearity across a wide range of starting material, revealing the same relative representation in cdna templates, regardless of gene abundance, making. The kit uses mmlv reverse transcriptase with lower rnase h activity, which avoids degradation of rna when rnadna hybrid forms in the progress of cdna synthesis. For the qscript cdna synthesis kit we recommend doubling the reaction volume to 40ul. Biorad iscript cdna synthesis kit invitrogen superscript iii firststrand synthesis supermix quanta qscript cdna synthesis kit. Rnase h which is component of this kit can be properly used if necessary. Reverse transcriptases rts use an rna template and a primer complementary to the rna to direct the synthesis of the first strand cdna, which can be used directly as a template for the polymerase chain reaction pcr. Does the edta have an influence on the synthesis of the cdna. Superscript vilo cdna synthesis kit catalog number 11754050 and 11754250 doc. The applied biosystems highcapacity cdna reverse transcription kit uses the random primer scheme for initiating cdna synthesis.
The superscript vilo cdna synthesis kit is designed to generate first strand cdna for twostep rtqpcr applications. Use the iscript cdna synthesis kit for firststrand cdna synthesis for realtime qpcr. First strand cdna synthesis reaction denaturation of rna and primer at 70c for 5 minutes can remove secondary structures that may impede long cdna synthesis. When stored under these conditions the kit components are stable for a minimum of one year after ship date.
The sensifast cdna synthesis kit provides a rapid and sensitive method for firststrand cdna synthesis, which displays excellent linearity across a wide range of starting material. Order biolines sensifast cdna synthesis kit on zageno. Biorads iscript cdna synthesis kit biocompare product. Efficiency kits varied in their ability to efficiently generate cdna from different rna inputs linearity of cdna synthesis kits with bactin 15 20 25 30 35 401. Script cdna synthesis kit contains all reagents required for first strand cdna synthesis in one box combining simple handling with high flexibility. Monsterscript reverse transcriptase is a thermostable reverse transcriptase that completely lacks rnase h activity. This kit includes just three tubes comprehensive of the reagents required for successful rtpcr. The tonkbio first chain cdna synthesis kit for rtpcr is optimized to efficiently synthesize first chain cdna from mrna or total rna templates. Reverse transcriptases rts use an rna template and a primer complementary. The iscript reverse transcription supermix for rtqpcr reaction setup for a single cdna synthesis reaction iscript rt supermix is a sensitive, fast, and convenient reagent for gene expression analysis using realtime reverse transcription quantitative pcr rtqpcr and standard rtpcr. This kit is powered by primescript reverse transcriptase primescript rt, which has exceptionally strong stranddisplacement activity. The genome core has been using the biorad iscript cdna synthesis kit for at least two years, and has found it to be the best method in terms of ease, reliability, and use for a variety of applications.