Plasmid DNA Extraction Kit--Q&A

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Q&A

Can the plasmid DNA extraction Kit be applied on Gram (+) bacteria?

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?
A	This system is mainly designed to extract plasmid DNA from Gram (-) bacteria such as E. coli. Gram (+) bacteria have thicker cell walls so the cell Lysis buffers provided in the kit do not lyse them readily. However, extraction of plasmid DNA from Gram (+) bacteria can still be achieved with additional treatment. After re-suspending the pelleted bacterial cells in the PD1 Buffer, add lysozyme to give a final concentration of 3 to 5 mg/ml. Incubate the suspension at 37˚C for 30-60 minutes (or for a shorter time when 5 mg/ml lysozyme is used). This treatment weakens the cell wall of the Gram (+) bacteria. Add PD2, and follow the rest of the protocol. For certain Gram (+) bacteria with thin cell walls, such as Lactobacillus, applying a double amount of PD1, PD2, and PD3 Buffer may be enough to lyse the cells. Yet, we still recommend treating Gram (+) bacteria with lysozyme to facilitate cell Lysis.
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?

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?