siRNA / miRNA gene silencing Human 501 Mel and SK Mel 28 FANCD2

Get tips on using MammoCult™ Human Medium Kit to perform 3D Cell Culture Media Human breast cancer MCF-7 cells-Mammospheres

Get tips on using EZ DNA Methylation kit to perform DNA methylation profiling Gene specific profiling - Human ovarian tissue MGMT

Get tips on using EZ DNA Methylation kit to perform DNA methylation profiling Gene specific profiling - Whole blood (human) FKBP5

Get tips on using MammoCult™ Human Medium Kit to perform 3D Cell Culture Media Human breast cancer MDA-MB-231 cells-Mammospheres

DNA isolation and purification is the first critical step in sample preparation that helps ensure optimal performance of downstream assays like PCR, microarrays, and sequencing. Failure in yielding high-quality DNA would be the major reason that DNA doesn't work for the downstream application. To circumvent this, one should follow the recommended storage conditions to minimize DNA degradation by nucleases and shouldn't overload the purification system.

Bacterial culture is a process of letting bacteria multiply in a controlled fashion (temperature, humidity, oxygen content or shaking), in a predetermined culture medium (antibiotic resistance to obtain homogenous clones). It is an important step, especially during cloning, as a single cell can be grown homogeneously (on semi-solid or in liquid conditions) to obtain colonies. As mentioned, bacteria can be cultured in broth cultures (Luria broth or LB) or Petri dishes (Agar plates). A specific antibiotic can be added to the broth or agar plates in order to grow bacteria which have the gene insert conferring its resistance to that antibiotic. Following points are necessary to consider for optimal growth conditions: 1. In general, most bacteria grow well at 37C, but there are some strains which require growth temperatures between 25-30C. 2. It is ideal in broth cultures to fill the flask to ⅓ or less of the total flask volume for optimal aerobic growth. 3. Shaking speeds between 140-180 rpm are appropriate to ensure aeration and that the cells are surrounded by fresh media, and do not settle.

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When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.

Get tips on using Human ANGPTL3 ELISA to perform ELISA Human - Angiopoietin-Like 3 (AngptL3)