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OneScript Plus cDNA Synthesis Kit
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OneScript Plus Reverse Transcriptase (PCR)

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Description

Description

Synthesize cDNA from complex RNA templates (i.e. secondary structures and high GC content) with abm?s OneScript ? Plus Reverse Transcriptase. Engineered to perform under high temperatures (45?C – 55?C), abm?s OneScript? Plus Reverse Transcriptase can synthesize full-length cDNA libraries from RNA templates up to 15kb in length. In addition, OneScript?Plus Reverse Transcriptase has outstanding proofreading ability due to the presence of a fidelity-enhancing subunit, thus making this RTase an excellent choice for whole genome sequencing.

Technical Specifications

Form

Enzyme supplied with 5X RT buffer

Concentration

200 U/ul

Shipping Condition

Dry Ice

Storage Condition

Store all components at -20 ?C.

Notes

This product is ISO-13485 certified.

Application

? Synthesizing cDNA from a ssRNA
? DNA primer extension
? Sequencing dsDNA
? Constructing cDNA library
? Constructing libraries for serial analysis for gene expression (SAGE)
? Synthesizing cDNA in rapid amplification of cDNA ends (3? & 5? RACE)
? Producing template for use in RT-PCR or real-time RT-PCR
? Labelling 3′-end of duplex DNA via end-filling reactions
? Generating probes for hybridization

Product FAQ

Q1: What is the lower limit and minimal amount of DNA that can be recovered?

Picogram levels of DNA can be recovered. The limitation is based on sensitivity of detection method.

Q2: How to process naked DNA stored in DNA/RNA Shield?

Use the standard protocol (add 2 or 5 volumes of DNA binding buffer depending on DNA size).

Q3: What to do if ethanol addition to the DNA Wash Buffer was omitted?

The DNA will be eluted off the column. Rebind samples using the appropriate amount of DNA Binding Buffer and wash the column with the properly prepared wash buffer.

Q4: What happens if more DNA was loaded on the columns than the stated maximum binding capacity?

Oversaturation of the column can result in total DNA loss due to clogging of silica matrix.

Q5: How many times can columns be reloaded?

We recommend no more than 5 times as binding efficiency might decrease.

Q6: What is the lower limit and minimal amount of DNA that can be recovered?

Picogram levels of DNA can be recovered. The limitation is based on sensitivity of detection method.

Q6: What is the minimum input volume of DNA sample?

Working with volumes below 50 ?l can result in decreased recovery. We recommend raising the starting volume to 100??l with water to ensure optimal binding conditions.

Citation

99 / 100
Bioz Stars

SMARCA4 loss is synthetic lethal with CDK4/6 inhibition in non-small cel…

Nat Commun – Published 4 Feb 2019

‘no. 15027865), 16.5 ?l PBS, 0.5 ?l 1% digitonin, 0.5 ?l 10% Tween-20, 5 ?l H2 O) and incubated at 37 ?C for 30 min in a thermomixer with 1000 RPM mixing. . After cleaning with Zymo DNA Clean and Concentrator-5 Kit (Zymo Research cat. no. D4014), eluted DNA was pre-amplified for 5 cycles using NEBNext 2x MasterMix (NEB cat. no. M0541).. Additional amplification cycles were added based on qPCR amplification .’

 

SLFN11 blocks stressed replication forks independently of ATR

Mol Cell – Published 01 Feb 2019

‘the transposase reaction mix [25 ?l 2x TD buffer, 2.5 ?l Transposase (Illumina) and 22.5 ?l of nuclease free water], and incubated for 30 minutes at 37 ?C. . Directly following transposition, the sample was purified using a DNA Clean and Concentrator-5 (ZYMO RESEARCH) and eluted with 25 ?l of DNA elution buffer.. Following purification, we amplified library fragments using 1x NEBnext PCR master mix and 1.25 ?M of custom Nextera PCR primers 1 and 2, using the ‘

 

TaME-seq: An efficient sequencing approach for characterisation of HPV g…

Sci Rep – Published 24 Jan 2019

‘Samples were subjected to tagmentation using Nextera DNA library prep kit (Illumina, Inc., San Diego, CA).. Tagmented DNA was purified using DNA Clean & Concentrator?-5 columns (Zymo Research, Irvine, CA) according the manufacturer?s instructions or ZR-96 DNA Clean & Concentrator?-5 plates (Zymo Research, Irvine, CA) according to the Nextera? DNA Library Prep Reference Guide (15027987 v01) before PCR amplification for target enrichment.. Amplification was performed using Qiagen Multiplex PCR Master mix (Qiagen, Hilden, Germany) according to the manufacturer?s instructions.’

See more details on Bioz

Reviews

?This kit performed as described. I was able to obtain good quality and yield of the DNA -better than with a couple of competitor’s kits.?

– Kathleen B. (SUNY-ESF)

?The simple steps and washing instructions were excellent. DNA was ready for use for PCR and gave strong results.?

– Tyler C.

?It only took 2 minutes for the total procedure – shorter than my current method of PCR purification.?

Marissa V. (Harvard Medical School)

?Almost no loss of elution buffer at the final step. It is quite impressive compared to the other kits I have used, which lose about 5-7?l.?

– Takashi K. (Monash University)

?I like the enhanced design of the column as it funnels every drop of volume directly through the silica in a very efficient manner. The quick spin times and relative ease of use are also a big draw as it cuts down on processing time and allows the attack of subsequent enzymatic steps at a heightened pace.?

– Joseph R. (Miller School of Medicine, University of Miami)

?I sampled this kit next to a kit we already use. I had three of the same sample, two for the Zymo kit and one for the other. On one of the ones designated for your kit, I accidentally added the sample directly to the column filter, then added other reagents (I figured I might as well try it). The other I did as per the instructions. The one on which I goofed gave a similar purification and concentration as the other kit, while the one I did properly yielded approximately four times the DNA per ?L as the other kit. I am very happy with this result, and when I rerun my samples, I plan to use your product.?

– Rosalind P. (University of Arkansas for Medical Sciences)

?The elution volume is low enough to concentrate any sample very well and the efficiency is not compromised at all.?

– Shanshan L. (UT Southwestern)

?It was a very simple procedure and it gave a concentration ten times the original amount.?

– Kimberly M. (University of Pennsylvania)

Kit Components

Cat #NameSize
D3004-4-10DNA Elution Buffer10 ml
D3004-4-4DNA Elution Buffer4 ml
D4003-1-LDNA Binding Buffer50 ml
D4003-2-24DNA Wash Buffer (Concentrate)24 ml
D4003-2-6DNA Wash Buffer (Concentrate)6 ml
C1001-50Collection Tubes50 Pack
C1001-500Collection Tubes500 Pack
C1001-1000Collection Tubes1000 Pack
C1008-50Zymo-Spin II Columns50 Pack
C1008-250Zymo-Spin II Columns5 x 50 Pack
D4004-1-LDNA Binding Buffer100 ml
C1078-50Zymo-Spin IICR Columns50 Pack
C1078-250Zymo-Spin IICR Columns5 x 50 Pack
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