Highlights
- Simple, rapid recovery of ultra-pure DNA from PCR, endonuclease digestions, and cell-free DNA preps, etc.
- Unique column construction allows sample loading and washing to be performed using a centrifuge, microcentrifuge, or vacuum source.
- Eluted DNA is well suited for use in PCR, DNA sequencing, DNA ligation, endonuclease digestion, RNA transcription, radiolabeling, arrays, etc.
Description
The DNA Clean & Concentrator-100 (DCC-100) is a PCR purification kit designed for the rapid desalting and purification of up to 100 ?g of high quality DNA from PCR, large format restriction endonuclease digestions, or cell-free lysates. Eluted DNA is suitable for nucleotide sequencing, array analysis, PCR, nucleotide blotting, restriction endonuclease digestion procedures, as well as many other downstream applications requiring high quality DNA. The entire DNA purification/concentration procedure typically takes less than 20 minutes and can be performed using a syringe, centrifuge or vacuum source together with a microcentrifuge.
Performance Data
Comparison to Finnzymes, ADAR
Comparison to Takara, ADAR
Comparison to SYBR GreenER, ADAR
Detergent Tolerance
?5% Triton X-100, ?5% Tween-20, ?5% Sarkosyl, ?0.1% SDS
Elution Volume
? 150 ?l
Equipment
Microcentrifuge and centrifuge or vacuum source.
Purity
Highly purified DNA is eluted with water and is especially well suited for sequencing, ligation reactions, and restriction endonuclease digestions.
Sample Source
Microcentrifuge
Purity
A260/A280 > 1.8, A260/A230 > 1.8
Sample Source
DNA from PCR, endonuclease digestions, DNA modification reactions, isotope/fluorescence labeling reactions, etc.
Size Range
50 bp to 23 kb
Yield
? 100 ?g total DNA can be recovered. For DNA 50 bp to 10 kb the recovery is 70-95%. For DNA 11 kb to 23 kb the recovery is 50-70%.
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.
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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.’
?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)
| Cat # | Name | Size |
|---|---|---|
| D3004-4-10 | DNA Elution Buffer | 10 ml |
| D3004-4-4 | DNA Elution Buffer | 4 ml |
| D4003-1-L | DNA Binding Buffer | 50 ml |
| D4003-2-24 | DNA Wash Buffer (Concentrate) | 24 ml |
| D4003-2-6 | DNA Wash Buffer (Concentrate) | 6 ml |
| C1001-50 | Collection Tubes | 50 Pack |
| C1001-500 | Collection Tubes | 500 Pack |
| C1001-1000 | Collection Tubes | 1000 Pack |
| C1008-50 | Zymo-Spin II Columns | 50 Pack |
| C1008-250 | Zymo-Spin II Columns | 5 x 50 Pack |
| D4004-1-L | DNA Binding Buffer | 100 ml |
| C1078-50 | Zymo-Spin IICR Columns | 50 Pack |
| C1078-250 | Zymo-Spin IICR Columns | 5 x 50 Pack |
| ROX | Low ROX | No ROX | Bio-Rad iCycler iQ systems |
|---|---|---|---|
| Applied Biosystems 5700 | Applied Biosystems 7500 | Quantabio Q | BioRad iCycler iQ? |
| Applied Biosystems 7000 | Applied Biosystems 7500 Fast | BioRad CFX | BioRad MyiQ? |
| Applied Biosystems 7300 | Stratagene Mx3000P? | Roche LightCycler 480 | BioRad iQ?5 |
| Applied Biosystems 7700 | Stratagene Mx3005P? | QIAGEN Rotor-Gene Q | |
| Applied Biosystems 7900 | Stratagene Mx4000? | Other | |
| Applied Biosystems 7900HT | Applied Biosystems ViiA 7 | ||
| Applied Biosystems 7900 HT Fast | Applied Biosystems QuantStudio? | ||
| Applied Biosystems StepOne? | Agilent AriaMx | ||
| Applied Biosystems StepOnePlus? | Douglas Scientific IntelliQube? |
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