Rna lysis buffer

Manufactured by Bio-Rad

Sourced in United States

The RNA lysis buffer is a reagent used in molecular biology techniques to isolate and purify RNA from biological samples. It functions by disrupting cell membranes and denaturing proteins, allowing for the release and stabilization of RNA molecules. The buffer is a crucial component in the RNA extraction and purification process.

Automatically generated - may contain errors

Lab products found in correlation

Paraformaldehyde (pfa), by Merck Group (2 mentions) Rneasy mini kit, by Qiagen (1 mentions) Iscript cdna synthesis kit, by Bio-Rad (1 mentions) Powerup sybr green master mix, by Thermo Fisher Scientific (1 mentions) Sybr green supermix, by Bio-Rad (1 mentions) Aurum total rna mini kit, by Bio-Rad (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Cfx96 system, by Bio-Rad (1 mentions) Penicillin streptomycin, by Merck Group (1 mentions)

Spelling variants of this product

Lysis buffer (41 citations)

4 protocols using rna lysis buffer

RNA Extraction and RT-qPCR Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fresh or frozen IECs were resuspended in RNA lysis buffer (Biorad). For total tissues, 1 cm pieces were homogenized in RNA lysis buffer (Biorad) or TRizol (Invitrogen). Accordingly, total RNA was extracted using Aurum Total RNA mini kit (Biorad) or using chloroform followed by RNeasy mini kit (Qiagen) purification. RNA was quantified by SpectraMax iD5 or NanoDrop. 500 ng of RNA was reversed transcribed using iScript cDNA synthesis kit (Biorad). Real-time PCR was performed using SYBR green supermix (Biorad) and detected on a CFX96 system (Biorad). For TNF^ΔARE samples, real-time PCR was performed using PowerUP SYBR green master mix (Applied Biosystems) on a QuantStuidio 6 (Applied Biosystems). Primers were ordered from Integrated DNA Technologies. Primer sequences are listed in Table 2. Relative expression was determined by 2^−dCt using Gapdh or Cdh1 as reference genes.

Lei X., Ketelut-Carneiro N., Shmuel-Galia L., Xu W., Wilson R., Vierbuchen T., Chen Y., Reboldi A., Kang J., Edelblum K.L., Ward D, & Fitzgerald K.A. (2022). Epithelial HNF4A shapes the intraepithelial lymphocyte compartment via direct regulation of immune signaling molecules. The Journal of Experimental Medicine, 219(8), e20212563.

+ Open protocol

+ Expand

Hypoxic Cell Culture Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Inflatable hypoxic chambers were adapted and modified from a published study^{47 (link)}, using sterile 4.5 mm heat-sealable pouches (VWR, USA). Briefly, culture dishes were placed inside pouches and the open end was heat-sealed to close. Two corners were then cut on a 45° angle just large enough to allow a 5 mL serological pipette fitted to flexible tubing (and attached to mixed air tank) to be inserted in one corner. The second corner was left open to allow air to exit the pouch as it was flushed with 1% oxygen mix. Pouches were flushed with 1% mixed air for approximately 3 minutes at which point the open corner was heat sealed. The pouch was then inflated to ~80% capacity at which point the remaining open corner was sealed and the chamber was placed in the tissue culture incubator. Individual chambers were made to allow for collection of cells after 2, 4, and 6 days and filled with mixed air containing 1% oxygen, 5% CO₂, and N₂ balanced (Airgas, USA). Cells were collected for RNA in RNA lysis buffer (Aurum, BioRad, USA), or were fixed in 4% paraformaldehyde (Sigma, USA), for 20 minutes at room temperature.

Natale B.V., Schweitzer C., Hughes M., Globisch M.A., Kotadia R., Tremblay E., Vu P., Cross J.C, & Natale D.R. (2017). Sca-1 identifies a trophoblast population with multipotent potential in the mid-gestation mouse placenta. Scientific Reports, 7, 5575.

+ Open protocol

+ Expand

Hypoxic Cell Culture Chambers

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Inflatable hypoxic chambers were adapted and modified as previously published^{15 (link),121 (link)}, using sterile 4.5 mm heat-sealable pouches (VWR, USA). Individual chambers were made to allow for collection of cells after 2, 4, and 6 days and filled with mixed air containing 1% oxygen, 5% CO₂, and N₂ balanced (Airgas, USA). Cells were collected for RNA in RNA lysis buffer (Aurum, BioRad, USA), or were fixed in 4% paraformaldehyde (Sigma, USA), for 20 minutes at room temperature.

Natale B.V., Mehta P., Vu P., Schweitzer C., Gustin K., Kotadia R, & Natale D.R. (2018). Reduced Uteroplacental Perfusion Pressure (RUPP) causes altered trophoblast differentiation and pericyte reduction in the mouse placenta labyrinth. Scientific Reports, 8, 17162.

+ Open protocol

+ Expand

Transcriptional Analysis of Perineal Tissue

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

For transcriptional analysis studies, samples of perineal or foreskin tissues were taken by punch biopsy on the first day, on day four, or day 20 after lesions first became visible on the perineum (5 animals per time point). Tissue samples from the perineum and foreskin of uninfected animals (N=5 animals) were collected in parallel for comparison. All samples were placed into 0.4 ml of Dulbecco’s Modified Eagle Medium (Corning Life Sciences-Mediatech, Inc., Manassas, VA) with 2.0% newborn calf serum (Life Technologies Incorporated, Carlsbad, CA) and 1.0% penicillin/streptomycin (Sigma-Aldrich, St. Louis, MO) and an equal volume of RNA lysis buffer (Bio-Rad, Hercules, CA) was added. Samples were frozen at −80°C prior to preparation of cDNA as described previously (Veselenak et al., 2018 (link)). Transcriptional analysis was performed as described previously (Veselenak et al., 2015 (link); Veselenak et al., 2018 (link)) for 44 immune-related genes and 4 housekeeping genes. New primers used in the current study and not described previously are listed in Table 4. Comparisons to identify significant changes in gene expression levels between tissue samples from uninfected and samples from each time point p.i. were calculated using Student’s t test with a p value cutoff of 0.05.

Bourne N., Banasik B.N., Perry C.L., Miller A.L., White M., Pyles R.B, & Milligan G.N. (2018). Development of disease and immunity at the genital epithelium following intrarectal inoculation of male guinea pigs with Herpes simplex virus type 2. Virology, 526, 180-188.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!