YALEPIC® Universal Genomic DNA Isolation Kit (No RNase A)

DNA yield varies by sample type and amount. Typically: 3-6 micrograms from 200 microL healthy whole blood; 10-30 micrograms from 25 mg animal tissue (e.g., liver); 15-40 micrograms from 5 x 10^6 cultured cells (e.g., HEK293). We recommend reviewing literature for target sample DNA content before experiments.

This is a universal kit with an optimized lysis buffer system suitable for various biological samples. Besides tissues, blood, and cells, it can effectively process bacteria (E. coli) and fungi (yeast). For plant tissues, FFPE (formalin-fixed paraffin-embedded) samples, and other special cases, it is not recommended.

DNA degradation relates to sample handling and operation. First, use freshly collected or properly frozen samples, avoiding repeated freeze-thaw (a major cause of nuclease-mediated DNA degradation). Second, insufficient lysis affects DNA integrity – ensure thorough tissue grinding and even mixing with lysis buffer. After extraction, aliquot DNA and store at -20 degrees C to avoid repeated freeze-thaw.

Common causes: Sample itself – low DNA content (low white blood cell count, over-cultured bacteria) or degradation from improper storage. Insufficient lysis – tissue pieces too large, proteinase K inactivated or insufficient incubation. Extend 56 degrees C incubation (overnight for difficult samples) and check proteinase K activity. Reagent issues – forgot to add ethanol to wash buffer (a common beginner mistake). Elution issues – pre-warm elution buffer to 60-70 degrees C, apply to membrane center, incubate 1-5 minutes before centrifugation, or perform double elution.

Purity is critical for downstream experiments: Low OD260/280 (below 1.9) indicates RNA residue. Add 4 microL RNase A (100 mg/mL) before proteinase K, incubate at room temperature for 5 minutes. DNA difficult to dissolve may indicate over-drying of precipitate. Air dry only, no more than 5 minutes, avoid vacuum drying.

Mainly due to inhibitor residues such as ethanol or salt ions in purified DNA. Solutions: spin empty column at max speed for 2-3 minutes after washing to thoroughly dry the column; ensure ethanol has been added to solutions as indicated on bottles. If DNA is impure, increase wash cycles or add BSA to PCR at 0.1 microgram/microL to mitigate inhibition.

Column clogging is typically caused by insufficient lysis or excessive sample input. Try: extend lysis time and increase inversion mixing; reduce starting sample amount; ensure lysate is completely clear before centrifugation.

The kit is very stable and can be stored and transported at room temperature (15-25 degrees C) with a shelf life of 18 months. Proteinase K (if in powder or solution form) should be aliquoted and frozen per protocol to avoid repeated freeze-thaw. No special cold storage needed, providing great convenience for daily laboratory use.