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Program 1: binding ( amp_ binding) program 2: washing and elution ( amp_ wash_ elute) deck position a: pcr 8 tube strip containing the ampure xp beads ( figure 1, blue), placed onto. pdf the liquid contains the end repaired dna. centrifugation with the lid open ensures that no ethanol remains during dna elution. incubate sample for 5 minutes at room temperature for maximum recovery. magnetic tube rack ampure xp beads lo- bind tubes freshly made 70% ethanol tips: you will get a better yield if you ampure xp beads protocol pdf increase the volume of ethanol for the washes enough to fill the tube.
protocol purify the pcr amplified cdna construct ( 100 μl) using a qiaquick pcr purification kit. 0x volume of ampure xp beads to the digested product from step 4 in a 1. it is recommended as part of the jetseqtm dna library preparation kit protocol for post ligation, post adapto. the result is a more purified pcr product.
it is powered by proven spri technology, which uses paramagnetic beads to selectively bind nucleic acids by type and size. introduction the agencourt ampure xp pcr1 purification systems utilize agencourt’ s solid- phase paramagnetic bead technology for high- throughput purification of pcr amplicons. 5 proteinase k working solution:. the assist plus pipetting robot immediately starts the protocol. com materials supplied by the user:. - for 30 μl of dna, add 54 μl of beads ( 30 x 1.
5b04023 spriselect vs ampure xp feature comparison spriselect and ampure xp can be substituted for each other in clean- up applications use the same sample to bead ratios are suggested for use by the most well- known library construction kit manufactures:. put ampure xp beads protocol pdf the mixture on a magnetic rack and let it sit for at least 2 minutes until the beads are concentrated against the magnet. find beckman coulter life sciences ampure xp technical documents here. to ensure the ampure xp buffer is homogenous, the beads are. remember, we are altering the standard ampure protocol by pdf leaving the dna bound to the beads while we end- repair, + a, and ligate adapters to dna fragments. agencourt ampure xp utilizes an optimized buffer to selectively bind pcr amplicons 100bp and larger to paramagnetic beads. high recovery of amplicons, greater than 100 bp. the amount of ampure® xp beads aliquoted to each well of the 8- tube strip depends on the number of columns in use and which wash step is being performed.
ampure xp bead transfer step: transferring ampure xp beads from an 8 tube pcr strip to a 96 well plate containing the dna samples. 8 μl of ampure xp beads per 1 μl of sample and let the mixture rest for 5 minutes at room temperature. add 110 μl of ampure beads to each sample, mix and rotate for 10 minutes at room temperature. the liquid should appear clear. how to: select and run the amp_ binding program on the voyager electronic pipette. such as data sheets, instructions for use ( ifus), certificate of analysis, and more. the assist plus automates many steps of a magnetic bead purification protocol and guides the pdf user through the remaining manual operations pdf to ensure an error- free process. note: changes that are part of the most recent revision are indicated in text by a bar in the margin of pdf the amended page. mix by pipetting or gently vortexing. the ampure xp bead- based reagent is intended for performing dna cleanup steps in different genomic applications like sequencing, qpcr/ ddpcr/ pcr, microarrays, and other enzymatic reactions. the end repaired dna is now bound to the beads.
have more questions? 1 prepare an 8- tube strip by adding well mixed, room- temperature ampure® xp beads ampure xp beads protocol pdf to each well of the strip in the volumes indicated in table 2 below. critical step: before adding shake well the agencourt ampure xp beads to resuspend any magnetic particles that may have settled. excess primers, nucleotides, salts, and enzymes can be removed using a simple washing procedure.
8 μl ampure xp beads ( beckman coulter) per 1. requiring no centrifugation or filtration, ampure xp reagents can be easily used in manual and automated 96- or 384- well formats. product ampure xp technical documents where can i find ampure xp technical documents? product no: a63881 the ampure xp reagent, part of our spri reagent family, is a highly efficient, easily automated pcr purification system that delivers superior quality dna with no salt carryover.
incubate the mixture for 5 minutes at room temperature. ampure® xp bead solution 20 mg/ ml proteinase k 50mm tris- hcl ( ph 8. maximizing recovery, consistency, and speed to facilitate the entire ngs workflow, ampure xp is optimized to meet the stringent needs of today’ s genomic applications and to minimize the risk of losing important genetic information. mix well by pipetting. agencourt ampure xp magnetic beads ( beckman coulter) are an efficient way to clean up samples for pcr, ngs, cloning and microarrays. 3 record ampure xp beads protocol pdf the reagent lot numbers in lims. original instructions revision history issue aa, 08/ agencourt ampure xp information for use version b37419aa issue ab, 08/ updates were made to the following sections: pcr purification questions and answers. agencourt® ampure® protocol 000601v024 page 3 of 9 for questions regarding this protocol, call technical support at agencourtagencourt bioscience corporation, ampure xp beads protocol pdf a beckman coulter company yycummings center, suite 2450 y beverly, massachusetts 01915 y www. 5) 100% etoh distilled water 3. you can contact us or read more about our genomic reagent solutions. 4 retrieve a new 96- well plate and label it with the run name followed by “ purified libraries”.
required equipment: vortex mixer with a foam tube holder attachment ( similar to vwr part#, or a rotator. note ampure xp is supported for snp 6. agencourt ampure xp utilizes an optimized buffer to selectively bind dna fragments 100 bp and larger to paramagnetic beads. the kit provides a solution for medium to high throughput requirements when carried out in a 96 well plate, but the protocol involves many washing and transfer steps that make it tedious to perform manually. place the samples on a magnetic separator, pdf when the beads have collected to the wall of the tube and the solution is clear, remove and discard the liquid. important: before eluting the dna from the column, centrifuge the column with the lid of the spin column open for 5 minutes at 13, 200 rpm.
genomic solutions cleanup and size selection ampure xp protocol ampure xp bead- based reagent protocol for pcr purification the protocol describes procedures of purification ( cleanup) of pcr products in well format using pdf the ampure xp reagent powered by our spri technology. this is the suggested protocol for clean- up using ampure xp beads.
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