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Molecular Biology and Genomics |
Molecular Biology and Genomics Articles
Q: Does AMPIPROBE® Bacterial Vaginosis Assay Kit, ENZ-GEN205-0100 include Megasphaera 1 as a control? Can this control be purchased as a stand alone item?
Viewed 82 times since Thu, Jun 10, 2021
ANS: Positive PCR BV control has Megasphaera as a kit component and they are part of the BV PCR kit ENZ-GEN205. The extraction BV Extraction control has Megasphaera, ENZ-GEN206 and is available as a stand-alone product. http://www.enzolifesciences... Read More
Is it possible to perform two-color detection by staining E6/E7 mRNA and labeling another protein with Alexa fluor 594 conjugated antibody with ENZ-300?
Viewed 49 times since Thu, Jul 29, 2021
We have not validated two-color detection by staining E6/E7 mRNA and labeling another protein with Alexa fluor 594 conjugated antibody. The E6/E7 RNAs are not labeled, they are hybridized with a labeled probe. The fixation step could cause some... Read More
How can RNA degradation in the sample be prevented?
Viewed 41 times since Wed, May 5, 2021
When RNA is the target of ISH, preservation of RNA in the sample is very important given the ubiquitous presence of RNase enzymes which can quickly degrade the sample. Gloves, sterile equipment, and RNase-free labware and reagents should be used at... Read More
What are the advantages of digoxigenin labeling?
Viewed 40 times since Fri, May 7, 2021
For chromogenic ISH, most commonly biotinylated probes are used. Digoxigenin offers the advantage of lower background, higher specificity, and the ability to combine it with biotin labeling in multiplexing, whether two different in situ probes are... Read More
Probe Design and Generation - How are DNA probes prepared?
Viewed 36 times since Wed, May 5, 2021
First choice is to explore if commercially ready-to-use probes are available for specific targets. Enzo Life Sciences offers a selection of ready-made, high-sensitivity PATHO-GENE® HPV or BIOPROBE® Infectious Agent Probes for HPV, Adenovirus,... Read More
How big are ISH probes+
Viewed 35 times since Fri, May 7, 2021
There is no defined answer. Depending on the tissue specimen, target type, and sequence, ISH probes might vary from below hundred base pairs (bp) to several hundred kbp. Most commonly, probes will be in between 200-1500bp for FFPE tissue sections.... Read More
How are ISH probes labeled?
Viewed 34 times since Fri, May 7, 2021
There are three main techniques for probe labeling. Labeled probes can be generated by incorporating labeled nucleotides during the synthesis or transcription of the DNA/RNA. Nick translation uses existing DNA templates. DNAse introduces single... Read More
How important are denaturation and hybridization temperatures?
Viewed 34 times since Fri, May 7, 2021
Correct denaturation and hybridization temperatures are paramount for successful ISH. Regardless of the probe or target being RNA or DNA, it is vital that the probe is properly denatured (normally at 95°C) just prior to application onto the... Read More
RNA Probe Design and Generation - How are RNA probes prepared?
Viewed 33 times since Wed, May 5, 2021
RNA probes are usually prepared with biotin or specific fluorophore by in vitro transcription. High specific activity RNA probes or riboprobes are synthesized from DNA templates cloned in expression vectors such as SP 6 and T 7 systems. Biotin... Read More
Why is peroxidase blocking necessary?
Viewed 32 times since Fri, May 7, 2021
When colorimetric detection by enzymatic reaction is used, a blocking step with peroxide solution is required to eliminate endogenous peroxidase activity of the tissue which can significantly decrease non specific background staining. Read More
What is the recommended ratio of dTTP to fluorophore-dUTP when using the Nick translation DNA labeling system 2.0?
Viewed 31 times since Thu, May 20, 2021
The SEEBRIGHT® fluorophore-labeled dUTPs are supplied as a 1 mM solution. We recommend to prepare working concentration of 0.3 mM by adding 10 µL of labeled dUTP to 23.3 µL of nuclease-free water. Store the diluted dUTP at -20˚C after use. For... Read More
Why are peptidases/proteinases used?
Viewed 30 times since Fri, May 7, 2021
Peptidases or proteinases like proteinase K, pepsin or trypsin are used in ISH to digest proteins contained in the sample that might mask the targeted DNA/RNA. Clearing proteins generally helps with probe penetration and peptidase/proteinase... Read More
What is the difference between ENZ-42910 and ENZ-GEN111
Viewed 28 times since Tue, Jun 8, 2021
The Nick Translation DNA Labeling System 2.0 (ENZ-GEN111) is an upgraded version of the old kit ENZ-42910. The new kit provides a simple, rapid and efficient method for generating labeled probes for FISH and CISH. A ready-to-use NT Enzyme Mix, that... Read More
What are ISH probes typically labeled with?
Viewed 27 times since Fri, May 7, 2021
Common labels include biotin, digoxigenin, and fluorochromes. Biotin is detected with streptavidin- conjugated enzymes followed by an enzymatic colorimetric reaction. Digoxigenin is detected with antibodies. Much like secondary antibodies in... Read More
Can ENZ-GEN300, FLOWSCRIPTTM HPV E6/E7 assay kit sort labeled cells by flow cytometry and then consider RNA seq analysis.
Viewed 26 times since Thu, Jul 29, 2021
We recommend Fixed and permeabilized cells hybridized with probes and analyzed by flow cytometry for E6/E7 transcript expression. We have not validated RNA seq analysis. Read More
How do we define Limit of Detection (LoD) and what is the percent reliability detection with AMPIPROBE® SARS-CoV-2 Assay Kit, ENZ-GEN215?
Viewed 25 times since Wed, Jun 16, 2021
The LoD is defined from this study as the lowest concentration of SARS-CoV-2 that can be reliably detected at least 95% of the time. Limit of Detection (LoD) of the AMPIPROBE® SARS-CoV-2 Test System for detection of SARS-CoV-2 N1 and N2 target ... Read More
Are the controls in AMPIPROBE® NG/CT/TV Extraction Controls, ENZ-GEN208-0500 quantitative or qualitative?
Viewed 23 times since Wed, May 5, 2021
Extraction controls are only qualitative and designed te used during genomic DNA extraction and is the qPCR assay set up to provide YES/NO answer in the qPCR Read More
What is the Limit of Detection (LoD) of the AMPIPROBE® SARS-CoV-2 Assay Kit, ENZ-GEN215 for detection of SARS-CoV-2 N1 and N2 target?
Viewed 22 times since Tue, Jun 15, 2021
The study results show that the LoD for the AMPIPROBE® SARS-CoV-2 Assay on samples isolated using the QIASymphony® SP instrument with the QIASymphony® DSP Virus/Pathogen Midi Kit is 280 copies/mL. LoD for the AMPIPROBE® SARS-CoV-2 Test System... Read More
Are there any cell damage or deterioration of RNA quality by performing a series of operations ENZ-GEN300, FLOWSCRIPT (TM) HPV E6/E7 assay kit?
Viewed 22 times since Thu, Jul 29, 2021
The loss or deterioration of the RNA is more likely to occur with the storage of the cells before analysis, although once the cells are out of the Thin-Prep medium, RNases are more likely to be active. The more operations done after the removal of... Read More
Does BIOARRAY™ Single-round RNA amplification and biotin labeling system, ENZ-42420 and ENZ-42421 have all the items necessary for amplification and labeling
Viewed 20 times since Thu, Aug 12, 2021
This kit contains all the buffers, enzymes, purification columns, elution tubes for cDNA synthesis from 250 ng up to 5000 ng total RNA. BIOARRAY™ Single-round RNA amplification and biotin labeling system is complete kit, composed of reagents for... Read More
Do we offer a kit to make cDNA from a low amount of RNA in the range 10-100 ng sample?
Viewed 20 times since Thu, Oct 7, 2021
For cDNA preparation from low input RNA samples, we offer AMPIGENE® cDNA Synthesis Kit, ENZ-KIT106. The AMPIGENE® cDNA Synthesis Kit uses the latest developments in reverse transcriptase technology and buffer chemistry to enhance cDNA synthesis... Read More
Does ENZO PATHO-GENE® HPV probes target E1, L2, E6 and E7 regions in the HPV genome? What is known about the targeting region of HPV probes?
Viewed 20 times since Wed, Mar 30, 2022
All of the PATHO-GENE® HPV probes we offer, vary in size from 100 bp to 1 Kb and the probes span long control region (LCR). Any additional information is proprietary. The specific recognition sequences are proprietary. Probes for specific... Read More
How was the Limit of Detection (LoD) determined for AMPIPROBE® SARS-CoV-2 kit, ENZ-GEN215?
Viewed 19 times since Wed, Jun 16, 2021
Preliminary LoD was determined testing triplicates of eight samples prepared by spiking the SeraCare AccuPlex™ SARS-CoV-2 Reference Material (Material Code 0505-0126, Batch #10480311, SARS-CoV-2 genomic RNA sequences ORF1a, RdRp, E gene, and N gene... Read More
What is the purity, volume and amounts of Bio-17-ATP, ENZ-42817 provided?
Viewed 19 times since Wed, Jun 1, 2022
Biotin-17-adenosine-5'-triphosphate is ≥93% (HPLC), the amount is 250 nmol in a volume of 25 ul. Read More
Can gene-specific primers be used for cDNA synthesis with BIOARRAY™ Single-round RNA amplification and biotin labeling system, ENZ-42420 and ENZ-42421?
Viewed 18 times since Thu, Aug 12, 2021
RNA amplification with BIOARRAY™ kit requires the use of primers included in the kit. RNA amplification with this kit requires the use of oligo dT primers and substituting with random or gene-specific primers with this kit is not recommended. Read More
What is the suggested volume of SEEBRIGHT™ Red 598 dUTP, ENZ-42855 and SEEBRIGHT™ Green 496 dUTP, ENZ-42831 with Nick translation DNA labeling system 2.0, ENZ-GEN111-0050?
Viewed 16 times since Thu, Oct 7, 2021
We suggest to reconstitute the dyes (0.3 mM) and use the same volumes as mentioned in the manual. A ratio of 1:2 volume of dye : dTTP for both the green and the red is suggested. https://www.enzolifesciences.com/fileadmin/files/manual/ENZ... Read More
How does PATHO-GENE® HPV Type 11 Probe, ENZ-GEN151 work?
Viewed 16 times since Wed, Mar 30, 2022
The ENZO PATHO-GENE® HPV Type 11 Probe is a biotin-labeled DNA probe designed to hybridize to HPV Type 11 DNA sequences. The probe is denatured and hybridized to the tissue section fixed on the microscope slide. Specific hybridization between... Read More
What is the recommendation to reconstitute SEEBRIGHT™ Red 598 dUTP, ENZ-42855 and SEEBRIGHT™ Green 496 dUTP, ENZ-42831?
Viewed 15 times since Thu, Oct 7, 2021
Dissolving in 10 mM TE pH8, is the preferred method since the nucleotides are stable for long term -20°C storage. When EDTA is not preferred, dissolving in nuclease-free water can be considered. Read More
What is the size of ENZO PATHO-GENE HPV probes and what regions do they target?
Viewed 15 times since Wed, Mar 30, 2022
All of the PATHO-GENE® HPV probe we offer, vary in size from 100 bp to 1 Kb and the probe span long control region (LCR). Any additional information is proprietary. Read More
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