We have updated the storage temperature of the purification columns for this product. For better long-term performance, it is recommended to store the purification columns at 2°C to 8°C.
My downstream enzymatic reactions are not working after purifying my DNA with your PureLink Quick Gel Extraction Kit. What should I do?
Here are some suggestions for your experiments:
- Many enzymes, including restriction endonucleases and ligases, are inhibited by small amounts of agarose and by perchlorate contaminants. This is not normally a problem. If it is, however, you can use the DNA without repurification by increasing the amount of enzyme used in the digest or ligation or by increasing the digestion incubation time. Also, you may use less DNA in your digest or ligation with the same total volume and the same amount of restriction enzyme.
- There may have been residual ethanol in the eluted fragment. Be sure to thoroughly centrifuge to remove the wash buffer, discard the wash buffer, and use a fresh tube to collect the eluted DNA. For applications that are very sensitive to ethanol, let the open spin column stand for 15 minutes at room temperature to let any excess ethanol evaporate.
- The washing steps may not be as efficient as they should be. Under these circumstances, there may be trace amounts of perchlorate in the eluate. To avoid this, extend the centrifugation times to 5 minutes and wash 2 times with wash buffer.
- Be sure to perform the optional wash step if you are using higher concentrations of agarose or are adding more than 250 mg to the cartridge. If applications are sensitive to EDTA, elute with water (pH 7.5-8.5), or with 10 mM Tris, pH 8.0 without EDTA.
What types and concentrations of agarose are compatible with the PureLink Quick Gel Extraction Kit (Cat. No. K210012)?
Both regular and low-melting agaroses can be used. When the agarose concentration is above 2%, use 600 µL of solubilization buffer for each 100 mg of gel.
Can I use water to elute my sample using the PureLink Quick Gel Extraction Kit?
Yes, water may be used, but please ensure that the water is clean and the pH of the water is between 7.5 and 8.5.
I'm running an agarose gel with TAE or TBE. Can I use your gel extraction kits?
Yes, both TAE and TBE agarose gels are compatible with our gel extraction kits.
A naturally occurring splice variant of CXCL12/stromal cell-derived factor 1 is a potent human immunodeficiency virus type 1 inhibitor with weak chemotaxis and cell survival activities.
Authors:Altenburg JD, Broxmeyer HE, Jin Q, Cooper S, Basu S, Alkhatib G,
Journal:J Virol
PubMed ID:17507482
CXCL12/stromal cell-derived factor 1 is a member of the CXC family of chemokines that plays an important role in hematopoiesis and signals through CXCR4 and CXCR7. Two splice variants of human CXCL12 (CXCL12alpha and CXCL12beta) induce chemotaxis of CXCR4(+) cells and inhibit X4 infection. Recent studies described four other novel ... More