The RNA was extracted from lymphocytes in MagNA Pure LC2.0 Automatic extractor with MagNA Pure LC Total Nucleic Acid Isolation Kit–High Performance: automatic RNA extraction using magnetic beads (Roche Diagnostics, IN, USA). The isolation protocol was performed according to the manufacturer's instructions. After extraction, RNA was quantified with a NanoDrop2000 (Thermo fisher Scientific, USA); Purity ranging between 1.8 and 2.1 was deemed as a good RNA quality.
Magna pure lc 2
Manufactured by Roche
Sourced in United States, Switzerland, Germany, Italy
The MagNA Pure LC 2.0 is a fully automated, high-throughput nucleic acid purification system. It utilizes magnetic bead-based technology to efficiently extract and purify DNA, RNA, and viral nucleic acids from a variety of sample types.
Automatically generated - may contain errors
Lab products found in correlation
Magna pure lc total nucleic acid isolation kit, by Roche (11 mentions)
Magna pure lc, by Roche (5 mentions)
Magna pure lc dna isolation kit, by Roche (3 mentions)
Magna pure compact, by Roche (3 mentions)
Dna isolation kit 1, by Roche (3 mentions)
Nanodrop 1000, by Thermo Fisher Scientific (2 mentions)
Qubit 2, by Thermo Fisher Scientific (2 mentions)
Thinprep media, by Hologic (2 mentions)
Lightcycler 96 system, by Roche (2 mentions)
Vacutainer k2edta tube, by BD (2 mentions)
Infinium methylationepic beadchip kit, by Illumina (2 mentions)
Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (2 mentions)
Primescript rt reagent kit, by Takara Bio (2 mentions)
Qiasymphony, by Qiagen (2 mentions)
Superscript 4 reverse transcriptase, by Thermo Fisher Scientific (2 mentions)
Altostar purification kit 1, by Altona Diagnostics (2 mentions)
Easymag, by bioMérieux (2 mentions)
Magna pure 96, by Roche (2 mentions)
Ribopure rna purification kit, by Thermo Fisher Scientific (2 mentions)
Total nucleic acid isolation kit, by Roche (2 mentions)
66 protocols using magna pure lc 2
1
Automated Lymphocyte RNA Extraction
2
Herpes Virus Detection in Autoimmune Encephalitis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Archived and frozen CSFs (2011–2015) from 77 patients with autoimmune encephalitis and known positivity for a neural antibody in one or more of the CBAs described above and 77 controls (antibody-negative CSFs from Mayo patients [2015] without known inflammatory or infectious CNS diseases) were tested retrospectively for herpes viruses (HSV-1/2, VZV, EBV, CMV, and HHV-6) by real-time PCR. The median patient age was 25 years (range, 2–85); 55 were female. Median control age was 49 years (range, 16–86); 49 were female. Control diagnoses were cognitive disorders (n = 33), epilepsy (n = 15), movement disorders (n = 11), no neurologic diagnosis (n = 17), or motor neuronopathy (n = 1).
For herpes virus real-time PCR testing, DNA from 200 μL of raw CSF was extracted on the MagNA Pure LC 2.0 (Roche Diagnostics, Indianapolis, IN) using the Total Nucleic Acid extraction protocol. Subsequently, 5 μL of extracted nucleic acid was added to a LightCycler capillary (Roche) containing 15 μL of master mix for one of the following: CMV (laboratory developed test [LDT]), EBV (Roche analyte specific reagents), HHV-6 (LDT), HSV-1 and -2 (Roche analyte specific reagents), and VZV (LDT). Testing was then performed on a LightCycler 2.0 (Roche) as previously described.17 (link)– (link)20 (link)
For herpes virus real-time PCR testing, DNA from 200 μL of raw CSF was extracted on the MagNA Pure LC 2.0 (Roche Diagnostics, Indianapolis, IN) using the Total Nucleic Acid extraction protocol. Subsequently, 5 μL of extracted nucleic acid was added to a LightCycler capillary (Roche) containing 15 μL of master mix for one of the following: CMV (laboratory developed test [LDT]), EBV (Roche analyte specific reagents), HHV-6 (LDT), HSV-1 and -2 (Roche analyte specific reagents), and VZV (LDT). Testing was then performed on a LightCycler 2.0 (Roche) as previously described.17 (link)– (link)20 (link)
3
Automated Nucleic Acid Extraction Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An automated MagNA Pure LC 2.0 instrument (Roche, Switzerland) was used for extraction of total nucleic acid (NA) or DNA. For the samples from Uganda as well as the parasitology samples from Mali, a MagNA Pure LC high-performance total nucleic acid isolation kit was used. For samples from Burkina Faso, Kenya, and the first season of the trial in the Gambia (both full blood in EDTA and saliva samples), MagNA Pure LV DNA isolation kits were used. The saliva samples collected after the second season of the trial in the Gambia were extracted using a Maxwell 16 instrument (Promega, USA) and Maxwell 16 DNA purification kits. Concentration measurements were done using a NanoDrop device (Thermo Fisher, USA) (only DNA from full blood in EDTA) and a Qubit fluorometer (Thermo Fisher, USA) with the Qubit HS (high-sensitivity) kit, which is specific for double-stranded DNA (dsDNA).
4
RNA Extraction from Nasopharyngeal Swabs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Nasopharyngeal swab specimens were collected in 2 ml of viral nucleic acid sample preservation fluid. Total RNA extraction was performed with the MagNA Pure LC Total Nucleic Acid Isolation Kit using the MagNa Pure LC 2.0 automated nucleic acid purifier (Roche), as recommended by the manufacturer.
5
Profiling Preterm Infant Ocular Fluids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Venous blood (0.5–1 mL) was collected from the ROP and no-ROP preterm babies by venipuncture. DNA was extracted from the blood samples using an automated DNA extraction platform (MagNa Pure LC 20, Roche) following the manufacturers guidelines. Likewise, for proteomic studies, the vitreous humor samples (100–500 µL) were collected from preterm babies with stage IV and V ROP (n = 30) who had undergone vitrectomy as a part of their routine clinical management. The controls for the proteomic studies included babies with congenital cataract (<6 months of age) who underwent partial vitrectomy as part of the surgical management (n = 30). The vitreous samples were immediately centrifuged at relative centrifugal force (rcf) of 10,621 g and the supernatant was stored at −80°C deep freezer until further use.
Additionally, crude tears were collected before instilling any drops or drug for pupil dilation in the eyes of preterm babies with ROP (Stage II–V) (n = 27) and no-ROP (n = 13) using a capillary tube and without touching the conjunctiva. The tear samples for ROP subjects were collected during the active disease condition either before or after 3 months following medical intervention.
Additionally, crude tears were collected before instilling any drops or drug for pupil dilation in the eyes of preterm babies with ROP (Stage II–V) (n = 27) and no-ROP (n = 13) using a capillary tube and without touching the conjunctiva. The tear samples for ROP subjects were collected during the active disease condition either before or after 3 months following medical intervention.
6
Genomic DNA Extraction and Verification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the samples from Populous collection, procedures concerning DNA extraction and sample processing have previously been described [48 (link),49 (link)].
For the group of psoriatic patients, genomic DNA was extracted from whole blood using the MagNa Pure LC 2.0 (Roche, Basel, Switzerland). NanoDrop 1000 (Thermo Fisher Scientific Inc., Waltham, MA, USA) and Qubit 2.0 (Thermo Fisher Scientific Inc., Waltham, MA, USA) were used to determine DNA quality and concentrations. Afterward, DNA samples underwent PCR reaction for sex verification [50 ].
For the group of psoriatic patients, genomic DNA was extracted from whole blood using the MagNa Pure LC 2.0 (Roche, Basel, Switzerland). NanoDrop 1000 (Thermo Fisher Scientific Inc., Waltham, MA, USA) and Qubit 2.0 (Thermo Fisher Scientific Inc., Waltham, MA, USA) were used to determine DNA quality and concentrations. Afterward, DNA samples underwent PCR reaction for sex verification [50 ].
7
HPV Genotyping in Cervical Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cervical exfoliated cells were collected in ThinPrep media (Hologic, Bedford, MA, USA) using a cytobrush from Japanese patients diagnosed as negative for intraepithelial lesion or malignancy (NILM), CIN1, CIN2, CIN3, or ICC at Keio University Hospital, Tsukuba University Hospital, and Showa University Hospital from 2012 to 2017. The total cellular DNA was extracted from the recovered cells on a MagNA Pure LC 2.0 (Roche Diagnostic, Indianapolis, IN, USA), and subjected to PCR with PGMY09/11 primers to amplify HPV L1 DNA, followed by reverse blot hybridization for HPV genotyping, as described previously [29 (link)]. The study protocol was approved by the Ethics Committees at each hospital and the National Institute of Infectious Diseases, and written informed consent for study participation was obtained from each participant.
8
Genotyping NEIL2 SNP rs804271 in BRCA2 Carriers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Single Nucleotide Polymorphism rs1466785, located in the NEIL2 gene is a cancer risk modifier for BRCA2 mutation carriers [4 (link)]. Imputation using 1000 Genomes data showed that there were several SNPs in strong linkage disequilibrium (LD) with rs1466785, the original SNP reported in Osorio et al. [4 (link)]. Of these, we considered rs804271 to be the best candidate, given that it showed the most significant associations and that there existed functional data supporting its putative role in cancer [19 (link)].
DNA was extracted from peripheral blood of FBOC patients or LCLs using MagNAPure LC 2.0 (Roche Diagnostics, Indianapolis, Indiana) following the manufacturer’s instructions. DNA quantification and quality were assessed by NanoDrop® (ND-1000 V3.7.1). A specific Taqman probe for rs804271 was used to genotype the presence/absence of the polymorphism among the sample collection. Allelic discrimination assays were conducted using the 7900HT Fast Real-Time PCR System (Applied Biosystems). Probe design for rs804271 is (G>T) instead of (C>A). Along the manuscript we refer to the variant as G>T.
DNA was extracted from peripheral blood of FBOC patients or LCLs using MagNAPure LC 2.0 (Roche Diagnostics, Indianapolis, Indiana) following the manufacturer’s instructions. DNA quantification and quality were assessed by NanoDrop® (ND-1000 V3.7.1). A specific Taqman probe for rs804271 was used to genotype the presence/absence of the polymorphism among the sample collection. Allelic discrimination assays were conducted using the 7900HT Fast Real-Time PCR System (Applied Biosystems). Probe design for rs804271 is (G>T) instead of (C>A). Along the manuscript we refer to the variant as G>T.
9
Quantification of PCV2 in Serum
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total nucleic acids were extracted from serum samples using a MagNA Pure LC total nucleic acid isolation kit (Roche Diagnostics GmbH, Mannheim, Germany) operated on a MagNA Pure LC 2.0 automatic nucleic acid extraction instrument (Roche Diagnostics GmbH, Mannheim, Germany), according to the manufacturer’s instructions. To quantify the PCV2 load in serum samples, quantitative real-time PCR assay was tested and performed with a LightCycler® 96 system (Roche Applied Science, Basel, Switzerland) as described Tsai et al. [18 (link)].
10
Genetic Variant Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Genomic DNA extraction was performed with a MagNA Pure LC Total Nucleic Acid Isolation Kit (Roche Diagnostics, Indianapolis, Indiana, USA) using Magna Pure LC 2.0® equipment from Roche. DNA samples were stored at −20 °C until analysis.
Genetic analysis was performed to assess four clinically relevant variants of the ABCB1 gene (rs3789243, rs1128503, rs2032582, and rs1045642), two variants of the CES1 gene (rs71647871 and rs2307243), one variant of the SLC15A1 gene (rs2297322), and one variant of the NEU2 gene (rs2233385). Allelic discrimination by real-time PCR was performed with specific primers and TaqMan® probes. Approximately 50 ng of genomic DNA was added to a master mix (Applied Biosystems, Foster City, CA, USA) that contained 200 nmol of primers and 40 nmol of probes for each polymorphism. Samples were placed in a 96-well plate and amplified as follows: pre-PCR read at 60 °C for 30 s, then 10 min at 95 °C, followed by 50 cycles of 95 °C for 15 s, and 60 °C for 1 min. Post-read analyses were performed at 60 °C for 30 s. All assays were carried out in a 7500 Fast PCR System (Applied Biosystems), and analyses were performed with 7500 Fast System SDS software.
Genetic analysis was performed to assess four clinically relevant variants of the ABCB1 gene (rs3789243, rs1128503, rs2032582, and rs1045642), two variants of the CES1 gene (rs71647871 and rs2307243), one variant of the SLC15A1 gene (rs2297322), and one variant of the NEU2 gene (rs2233385). Allelic discrimination by real-time PCR was performed with specific primers and TaqMan® probes. Approximately 50 ng of genomic DNA was added to a master mix (Applied Biosystems, Foster City, CA, USA) that contained 200 nmol of primers and 40 nmol of probes for each polymorphism. Samples were placed in a 96-well plate and amplified as follows: pre-PCR read at 60 °C for 30 s, then 10 min at 95 °C, followed by 50 cycles of 95 °C for 15 s, and 60 °C for 1 min. Post-read analyses were performed at 60 °C for 30 s. All assays were carried out in a 7500 Fast PCR System (Applied Biosystems), and analyses were performed with 7500 Fast System SDS software.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!