Plasmid DNA Extraction Kit--Q&A

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Q&A

Why was Restriction enzyme digestion unsuccessful?

Plasmid DNA Extraction Kit(PD004, PD100, PD300) --Q&A
　

Q	Why do we sometimes obtain a lower yield?
Q	Why doesn’t the Eluted DNA perform well in downstream applications?
Q	Can the plasmid DNA extraction Kit be applied on Gram (+) bacteria?
Q	Why do we get eluted plasmid contaminated with RNA?
Q	Why can’t the eluted plasmid be cut by restriction enzymes properly?
Q	Why does the Plasmid DNA Extraction Kit require two washings with two types of wash buffers (W1 and Wash Buffer) while competitors kits require one washing?
Q	Eluted plasmid appeared degraded after storage for an extended period of time.
Q	Why was Restriction enzyme digestion unsuccessful?
A	There are a few possible reasons that could lead to failure in restriction enzyme performance of extracted plasmid. (1) Enzyme Activity: Most restriction enzymes are temperature sensitive. Prolonged storage or usage past expiry date will decrease enzyme activity dramatically, hence causing partial or total failure of plasmid cutting. (2) Sensitivity: Different strains of E. Coli will have varying sensitivity to dam or dcm Methylation. If digestion sites were blocked by overlapping dam or dcm Methylation during transformation, it will cause difficulty at the restriction enzyme digestion step. (3) Plasmid Concentration: High concentrations of final extracted plasmid will cause similar problems.

Genomic DNA Extraction Kit(GT004,GT050,GT300,GB004,GB100,GB300,GP004,GP100)--Q&A
　

Q	What should be done if the Column is clogged?
Q	Why is the DNA recovery lower than expected?
Q	Why doesn’t the Eluted DNA perform well in downstream applications?
Q	Why does the genomic DNA band appear smeared in agarose gel electrophoresis?
Q	What is the key to successfully isolating genomic DNA of good yield and quality?
Q	Why does the sample appear viscous after sample Lysis and not pass through the column easily, even when less than the suggested maximum sample amount is used?
Q	When Buffy coat is used, the column tends to be clogged.

Gel/PCR DNA Fragments Extraction Kit (DF004, DF100, DF300)--Q&A

Q	Can the Gel/PCR DNA Kit completely remove primers or dimer products from PCR reactions?
Q	Can the Gel/PCR DNA Fragments Extraction Kit be used to clean up DIG-labeled DNA fragment?
Q	A band, smaller than the desired DNA fragment is present in the eluant.
Q	Why DNA recovery is lower than expected?
Q	Why doesn’t the Eluted DNA perform well in downstream applications?

RNA Related Products(RT050,RT100,RT300,RB050,RB100,RB300,RP050,RP100)--Q&A

Q	How do I increase yields of total RNA?
Q	Why does the obtained RNA appear smeared and degraded?
Q	Is total RNA isolated by the Total RNA Extraction System free of genomic DNA?
Q	What is the most suitable kit for Cytoplasmic RNA extraction?
Q	What is the function of the lysate filter columns?
Q	Which kit is suitable for RNA extraction from yeast?