Get tips on using pX330-U6-Chimeric_BB-CBh-hSpCas9 to perform CRISPR Mouse - Deletion RAW 264.7 Nfatc1
Get tips on using pX330-U6-Chimeric_BB-CBh-hSpCas9 to perform CRISPR Mouse - Deletion RAW 264.7 Dcstamp
Get tips on using pX330-U6-Chimeric_BB-CBh-hSpCas9 to perform CRISPR Mouse - Deletion RAW 264.7 Nrp2
Get tips on using pX330-U6-Chimeric_BB-CBh-hSpCas9 to perform CRISPR Mouse - Deletion 3T3-L1 MmP13
Get tips on using MACSprep™ Chimerism CD34 MicroBead Kit, human to perform Cell Isolation CD34+ cells
Get tips on using pX330-U6-Chimeric_BB-CBh-hSpCas9 to perform CRISPR Mouse - Deletion ES (embryonic stem) cells Slx2
Get tips on using pX330-U6-Chimeric_BB-CBh-hSpCas9 to perform CRISPR Rat - Deletion INS-1 832/13 Ep300
Get tips on using pET-28a-chBCO2 to perform Protein Expression Prokaryotic cells - E. coli chicken BCO2
Get tips on using pBAD-Thio-TRP36 to perform Protein Expression Prokaryotic cells - E. coli TRP36 Ehrlichia canis
RNA quantification for appropriate concentration and quality (260/280 ratio) is an important step before downstream analysis (including sequencing, RT-qPCR, etc.). Having insufficient RNA quantities or a high salt or phenol in the RNA product can lead to variable or irreproducible downstream results. The various methods used for RNA quantification include: 1. UV spectrophotometric (challenges include: low sensitivity, cannot distinguish between nucleic acid species), 2. Fluorescence-based (challenges include: requires standards, cannot measure amplifiability, not sequence-specific), and 3. RT-PCR (challenges include: requires standards, time-intensive, costly). In order to overcome these challenges, and also to ensure the proper quantification and quality control for RNA product, it is important to use at least two or more methods in order to discard any inconsistencies. Using standards for calibrations increases the sensitivity range for RNA detention (fluorescence- and RT-PCR-based methods). When using RT- PCR, it is important to choose correct primers, aligning to the desired site on the template and of appropriate product length, along with positive, negative and loading controls. It is also important to have at least two primer pairs in order to confirm results.
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