Cat. No. / ID: 9001525
Cat. No. / ID: 9001514
|Sample Size||100-200 µl PCR samples|
|Elution volume||30-100 µl in increments of 10 µl, default 50µl|
|Starting material||Amplification reactions|
This protocol has been adapted by the North Louisiana Criminalistics Laboratory from the pretreatment for epithelial cells mixed with sperm cells and is for the lysis and extraction of DNA from a sperm pellet after differential separation.
The PyroMark Q48 Autoprep protocol uses magnetic streptavidin-coated Sepharose® beads (PyroMark Q48 Magnetic Beads), which bind to the biotinylated PCR strand.
PyroMark Q48 Magnetic Beads can be ordered at QIAGEN with the catalog no 974203.
Part numbers and prices for QIAcube accessories can be found by clicking here.
If you need to purchase a new computer for your PyroMark Q96, or if you need to reinstall, upgrade, or replace the operating system (OS), you can reinstall the PyroMark Q96 software without purchasing a new license.
PyroMark Q24 takes very little bench space, measuring only H420 x W390 x D525 mm.
Yes. The heated shaker on the QIAcube can be operated independently if the QIAcube is not performing a protocol run. For instructions on how to operate the shaker, refer to the QIAcube User Manual, chapter 5 'Operating Procedures, section 5.7 'Operating the shaker'.
The shaker can be used to heat and shake samples simultaneously, shake samples without heating, or heat samples without shaking.
See: http://www.qiagen.com/products/catalogpagecontrols/header/qiacube/360/default.aspx for a virtual demonstration of the heater and shaker on the QIAcube.
When looking at technical literature, pulse time settings, and other information, the following table can be used to determine the QIAGEN names for former Biotage instruments.
|PSQ 96||PyroMark Q96 ID|
|PSQ 96 MA||PyroMark Q96 ID|
|PSQ HS 96||PyroMark Q96 MD|
|PSQ HS 96A||PyroMark Q96 MD Automated|
An instrument method or instrument code encodes the individual pulse time settings of specific cartridge lot batch. These pulse time settings change when e.g. a new batch of capillaries is used with slight variations in the needle diameter. For larger diameters, the pulse settings are lowered to dispense the correct volume of liquid. In addition, the viscosity of enzyme and substrate mixes can change which influences dispensing volumes.
The individual instrument method/code number is printed on the cartridge label. The corresponding methods/code settings can be downloaded as a file from the respective instrument webpage and opened in the PyroMark application software.
Visit the QIAcube Protocols page to search for protocols. The instructions for transferring a protocol to the QIAcube are listed below.
To transfer new protocols to the QIAcube, first save them to the supplied USB stick.
Note: Protocols already installed on the QIAcube that have the same name as downloaded protocols will be overwritten.
8. When the protocols have been successfully transferred to the QIAcube, press “OK”.
Data generated with the methylation analysis software (detection and quantification of CpG sites) on PyroMark Q24 contains unique features that act as quality control for complete bisulfite conversion of DNA. When the assay encounters a C not followed by a G (C unmethylated), that C should be fully converted to T if the bisulfite treatment upfront was successful. Subsequently, it should be presented in the pyrogram as T=100%. This acts as a useful quality control for full conversion of unmethylated C residues during bisulfite treatment and PCR.
Yes. At the end of each protocol run on the QIAcube, a report file is generated that contains important information such as number of samples, liquid volumes measured during the load check, and run time. A log file is also generated that contains information about instrument actions, such as movement of the robotic arm.
This software will replace the old PyroMark Q96 ID software version 1.0. The PyroMark Q96 ID software version 2.5 contains four assay setup and analysis modes which are AQ, CpG, SNP and SQA. It is based on the Pyromark Q24 application software and is a file based software (runs are not stored in an ORACLE database). It replaces the PyroMark CpG software since a CpG mode is included. There is no SNP multiplexing support and no alignment functionality (as found in software version 1.0).
Yes. The QIAcube centrifuge can be operated independently if the QIAcube is not performing a protocol run. For instructions on how to operate the centrifuge, refer to the QIAcube User Manual, chapter 5 'Operating Procedures, section 5.8 'Operating the Centrifuge'.
Yes, this is absolutely necessary. If the lids are not securely fixed in the rotor adapter slots during processing on the QIAcube, they will become detached from the elution tubes and columns during centrifugation and will collect at the bottom of the centrifuge chamber.
PyroMark Q24: The mean single peak height is 95 +/- 55 RLU
PyroMark Q48: The mean single peak height is 70 +/- 40 RLU
PyroMark Q96 ID: The mean single peak height is 35 +/- 10 RLU
Pyromark Q96 MD: The mean single peak height should be at least 350 RLU
The following disinfectants and detergents are recommended for cleaning the QIAcube:
Lysetol® AF (Gigsept® Instru AF) (Schülke & Mayr GmbH) — disinfectant for submerging worktable items (consists of 14 g cocospropylene-diamine-guanidine diacetate, 35 g phenoxypropanols, and 2.5 g benzalkonium chloride per 100 g Lysetol AF, with anticorrosion components, fragrance, and 15–30% nonionic surfactants).
DECON-QUAT® 100 (Veltek Associates, Inc.) — quaternary ammonium salt based disinfectant concentrate for submerging worktable items (contains 5% alkyldimethylbenzylammonium chloride and 5% alkyldimethylethylbenzylammonium chloride).
Note: If you want to use disinfectants different from those recommended, ensure that their compositions are similar to those described above.
Up to 12 samples can be processed in a single run.
No. The QIAcube is designed for fully automated processing of up to 12 samples using QIAGEN spin-column kits. Do not use spin columns manufactured by other suppliers with the QIAcube. Damage caused by use of other types of spin columns will void the instrument warranty.
See: http://www.qiagen.com/products/catalogpagecontrols/header/qiacube/360/default.aspx for a demonstration on how the QIAcube is used with the RNeasy Mini Kit for automated purification of RNA.
No. Modified QIAGEN protocols or custom protocols developed to meet your specific requirements can be requested and purchased from QIAGEN.
1. Go to www.qiagen.com/MyQIAcube and enter the required information. If our Application Specialists require additional information, they will contact you directly.
2. You will be informed by e-mail when your customized protocol is ready for download.
3. Click the link in the e-mail to go to the download page.
See the PyroMark Gold Q96 Reagents Handbook for additional information.
PyroMark instruments offer a range of throughput scales. The PyroMark Q24 can process 1–24 samples in parallel, the PyroMark Q48 Autoprep 1-48, the PyroMark Q96 ID 1–96, and the PyroMark Q96 MD 1-96, or the automation option enables automated processing of ten 96-well plates. The sample processing speed depends on the number of nucleotide dispensations necessary for the programmed analysis. Twenty dispensations take approximately 24 minutes on all instruments; thus, 96 samples are typically processed in 10-100 minutes.
If you need to install the PyroMark Q24 software on a new computer replacing your old one, or after you reinstall or upgrade the operating system, you can reinstall the PyroMark Q24 software without purchasing a new license.
FAQ ID - 3473
Usually the sequencing primer is used at 0.3µM in annealing buffer but some assays might require additional optimization of the sequencing primer concentration.
The PyroMark Q48 Autoprep dispenses the sequencing primers for annealing. The final concentration of sequencing primers in a well is 0.8µM, but may be adapted to optimize assays.
See: http://www.qiagen.com/Products/Catalog/Automated-Solutions/Sample-Prep/QIAcube#orderinginformation for a virtual demonstration on how to use the QIAcube.
The QIAcube has the following weight and dimensions:
Please refer to section 6.5 'Decontaminating the QIAcube' in chapter 6 'Maintenance Procedures' of the QIAcube User Manual.
PyroMark Q24 analyses up to 24 individual samples in a single run. The time needed to perform a run depends on the number of nucleotides dispensed in the specific assay. A typical SNP assay contains 15 dispensations and a typical CpG assay 45. Since the estimated dispensation time is 1 minute/nucleotide, a typical SNP run takes 15 minutes to perform and a typical CpG assay takes 45 minutes.
You can stop a QIAcube protocol if there is an emergency by pressing “Cancel”. To confirm that you want to stop the protocol run, press “OK”. To cancel the 'stop protocol' command, press “Cancel”.
Note: If a protocol run is stopped, the run cannot be restarted. The samples must be processed manually. The last protocol step performed by the QIAcube will be displayed in the touch screen. For more information, refer to the QIAcube User Manual.
The corresponding manual kits can be used for all available protocols on the QIAcube. The manual kits contain sufficient buffers and reagents to process the number of spin columns provided with the kit.
In addition to the purification kit, Rotor Adapters (10 x 24), cat. no 990394, and the required disposable tips:
need to be ordered separately. To find out which filter-tips are required for a specific application, refer to the corresponding protocol sheet.
Typical reading length using Pyrosequencing technology is 40-60 bases. However, as with any sequencing technology, the maximum read length will depend on template secondary structure, base content, quality of PCR-product, and other parameters.
Depending on the sequence to be analyzed, highly accurate read lengths of 140 or more bases can be obtained in just a single reaction with the Q48 PyroMark Autoprep.
No. The QIAcube is equipped with an optical sensor, which determines the tip size and the number of tips loaded onto the worktable prior to starting a run.
The QIAcube can use partially used tip racks from previous runs, provided that the racks contain sufficient tips to process the number of samples loaded for the current run.
PyroMark Q24 uses Pyrosequencing technology for mutation analysis and provides a built-in quality control in each run. By sequencing nucleotides flanking the mutation of interest, the researcher gets confirmation that the assay was made at the correct position. In addition, blank dispensations are included in the assay providing a negative control of the run.