User:Jarle Pahr/Kits

Commercial kits for applications in molecular biology:

HisExpress protein purification columns: http://www.claremontbio.com/HisExpress_Protein_Purification_Columns_s/72.htm (available at http://www.bioquote.com/product.php?id=291680)

PrepEase his-purification: http://www.affymetrix.com/estore/browse/brand/usb/product.jsp?navMode=34000&productId=131366&navAction=jump&aId=productsNav#1_1

Affinity chromatography products at Jena Bioscience: http://www.jenabioscience.com/cms/en/1/browse/1450_affinity_chromatography.html?gclid=CJCPmK6HvrUCFQtY3godFWEAtg

Kit	Use	Advantages	Disadvantages	Time	Experience
Promega Wizard SV miniprep	DNA isolation	Quick	Low yield	45 - 60 min
Promega PureYield™ Plasmid Miniprep System	High-purity DNA isolation
Machery-Nagel NucleoBond Xtra Midi (midiprep)	DNA isolation	High yield (~20 ug), pure product.	Slow	2 d	Yields of 100-140 ng/uL in 200 uL, for a total of about 20 ug. Yield may depend on skill in handling washing and resuspension of DNA pellet. Might be more efficient to do several (~10) parallel minipreps instead?
Qiaquick gel extraction	DNA extraction from gel	Fast	Impure product		Impurities give absorbption peak at 230 nm in eluted product.
Qiaquick PCR purification	Purification of PCR products	Fast	15 min
QIAEX II gel extraction	DNA extraction from gel	Higher purity than qiaquick	Slower than Quiaquick	1-5 h	Yield of about 200 ng from ~1.5 ug starting amount of plasmid.
QIAGEN PCR Cloningplus Kit	Subcloning (TA cloning)	?	?	?
Dongsheng Bio Eco-friendly Plasmid miniprep kit	DNA isolation			?	-
Dongsheng Bio PCR and DNA Fragment Purification Kit	Purification of PCR fragments			?
QiaAmp

Notes:

Promega SV Wizard miniprep:

Observed plasmid yields:

Strain:Plasmid	Date	Culture volume incubated	Culture volume centrifuged	Medium /Antibiotic	Incubation vessel	Incubation time	OD600	Elution volume (uL)	Obtained DNA concentration (ng/uL)	Total yield (ug)	Yield (ug / mL * OD600)
Dh5a:pSB-M1g	21.5.13	10 mL	~4.5	LB + Kan 50 ug/mL	50 mL tube	~ 8h	~2.2	50	49	2,45	0,25
Dh5a:pSB-M1g	21.5.13	10 mL	~4.5	LB + Kan 50 ug/mL	50 mL tube	~ 8h	~2.2	50	35	1,75	0,18

Qiaquick gel extraction kit:

Qiaex II: Use a thermomixer (high mixing, RPM ~ 500 ) to get constant mixing during incubation.

In vitro transcription kits:

https://www.neb.com/products/e2050-t7-quick-high-yield-rna-synthesis-kit