FAQ MolYsis
Interesting Publications
Gyarmati et al. (2015): A study with 130 blood samples from hematological patients receiving dose-intensive antitumoral treatment were analyzed using MolYsis™ Complete5 and NGS sequencing. Link Thoendel et al. (2016): Previously negative sonicate ...
Which applications are possible with the eluted DNA (after MolYsis™ technology)?
The eluted DNA can be used for diverse analyses For example: broad-range PCR and Sanger sequencing, specific PCR assays, hybridization assays, multiplex PCR, panel systems, custom PCR assay, NGS metagenomic/shotgun sequencing, whole genome ...
What is the sensitivity of detection of bacteria and fungi in blood?
Sensitivity of detection / Bacteria & Fungi Spiking experiments showed that bacteria and fungi can be detected at low titers in whole blood using real-time PCR and subsequent Sanger Sequencing. For instance, with 5 ml whole blood using MolYsis™ ...
How can the eluted DNA concentration be measured?
Diverse approaches are possible, but please be aware that due to the depletion of human/animal DNA, the DNA concentration might be quite low (between ng and fg amounts of DNA are isolated depending on the sample used). Possible methods include: qPCR ...
Can I change the elution volume?
For all manual RUO kits (MolYsis™ Basic5, MolYsis™ Complete5, Ultra-Deep Microbiome Prep, and Ultra-Deep Microbiome Prep10) the elution volume might be changed. A change of the elution volume is not possible with the automated MolYsis-SelectNA™plus ...
Is the MolYsis™ technology applicable to plant samples?
The MolYsis™ technology has not been systematically tested for plant samples and, taking the structural properties of plant cells into account, the successful host DNA depletion of the plant DNA does not seem likely.
Is the MolYsis™ technology applicable for viruses/viral NA isolation?
The MolYsis™ technology has not been systematically tested for the isolation of viral NA.
After host DNA depletion with the MolYsis™ technology, can I isolate RNA?
RNase Please note that the MolYsis™-based kits are not systematically tested for the presence of RNases. After MolYsis™ Basic5 or step 7 of the other MolYsis™-based kits’ protocols, RNA isolation protocols could be followed.
Can blood cultures be used for the isolation of microbial DNA?
Blood culture All kits based on the MolYsis™ technology can be used for the isolation of microbial DNA from blood cultures. The blood culture samples shall be diluted at least 1:10 (100 µl blood culture + 900 µl SU buffer) before starting the ...
Can FFPE samples be analyzed with MolYsis™-based kits?
FFPE Sample Yes, FFPE samples can be processed by simply following the tissue protocol of the Ultra-Deep Microbiome Prep and Ultra-Deep Microbiome Prep10 kits. There are no additional steps like deparaffinization required.
What are the sample requirements? How shall samples be stored after sample collection? How fresh or old can samples be?
Please note: For any MolYsis™-based kits the microbial cells need to be intact! After taking the samples, please cool the samples in the fridge at ~8°C for short-term storage. Optimally, fresh samples are used for the highest detection sensitivity. ...
What is the sample volume that can be processed?
Please refer to the section “What are the differences between the kits including the MolYsis™ technology?“.
What kind of samples can be processed with MolYsis™?
MolYsis™ has proven valuable for the analysis of a wide range of sample materials including various tissue specimens, body fluids, and swabs, especially for applications in the field of NGS and PCR based detection methods. Samples evaluated for ...
What to do when there is no centrifuge in my lab that can reach 9,500xg for 50 ml tubes (related to MolYsis™ Basic5, MolYsis™ Complete5, or Ultra-Deep Microbiome Prep10 kit)?
No centrifuge for 9500xg/50ml tubes If your lab is not equipped with a centrifuge that can reach 9500xg for 50ml tubes*, you can use a lower speed (e.g., 4,500xg) and should extend the centrifugation time to 20 min instead of the 10 min in the ...
What to do when there is no thermomixer in my lab for the MolYsis™-based kits?
MolYsis & Thermomixer The MolYsis™ technology requires certain temperatures, 37°C, 56°C, and 70°C. During the incubation, samples should be mixed well. No Thermomixer? If no thermomixer with a shaking function is available, a heating block could be ...
What kind of consumables are to be used?
DNA-free pipette tips with filters are obligated to avoid carryover contamination during DNA preparation. Also, DNA-free 1.5 and 2.0 ml polypropylene tubes, as long as not supplied with MolYsis™ kits, should be used. Many manufacturers of plastic ...
Do I need to change my validated DNA purification system?
Not necessarily. Thanks to its modular nature, MolYsis™ Basic5 is adaptable to any DNA purification system. The kit is designed to treat <1 ml and 5 ml fluid samples including host DNA depletion and lysis of bacterial and fungal cells. Afterward, the ...
How is human DNA depleted during sample processing?
Human DNA depletion Samples are treated with a buffer lysing the human/animal cells, while bacterial and fungal cells stay intact. The released human/animal DNA is enzymatically degraded along with all other free DNA and bacterial and fungal cells ...
What are the differences between the kits including the MolYsis™ technology?
Please refer to the table below to see all differences: Kit name / specifications Sample materials & volumes Processing Host DNA depletion & lysis of bacterial and fungal cells Bacterial and Fungal DNA isolation MolYsis™ Basic5 Fluids < 1ml and 5 ml ...
Why deplete host (human or animal) DNA before PCR-based assays for bacteria and fungi?
Why is MolYsis useful? A major problem of the direct PCR-based detection of bacteria and fungi from clinical specimens is that host DNA can exceed microbial target sequences by several thousand-fold. Host DNA contains unspecific binding sites e.g., ...
What is MolYsis™?
MolYsis™ is a technology that lyses host (human or animal) cells and quantitatively depletes host DNA (and all other free DNA) by a DNase treatment. In the second step, intact microorganisms are lysed, and highly enriched microbial DNA is isolated. ...