User:Jarle Pahr/Kits: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Jarle Pahr (talk | contribs) No edit summary |
Jarle Pahr (talk | contribs) No edit summary |
||
Line 42: | Line 42: | ||
Promega SV Wizard miniprep: | Promega SV Wizard miniprep: | ||
Observed plasmid yields: | |||
{| class="wikitable" border="1" cellpadding="5" cellspacing="0" | |||
|- | |||
! Plasmid!! Date !! Culture volume !! Antibiotic concentration !! Incubation vessel !! Incubation time | |||
|- | |||
|pSB-M1g|| ||Culture volume || Kan 50 ug/mL|| 100 mL EM||? | |||
|} | |||
Qiaquick gel extraction kit: | Qiaquick gel extraction kit: | ||
Qiaex II: Use a thermomixer (high mixing, RPM ~ 500 ) to get constant mixing during incubation. | Qiaex II: Use a thermomixer (high mixing, RPM ~ 500 ) to get constant mixing during incubation. |
Revision as of 09:06, 9 May 2013
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:
Plasmid | Date | Culture volume | Antibiotic concentration | Incubation vessel | Incubation time |
---|---|---|---|---|---|
pSB-M1g | Culture volume | Kan 50 ug/mL | 100 mL EM | ? |
Qiaquick gel extraction kit:
Qiaex II: Use a thermomixer (high mixing, RPM ~ 500 ) to get constant mixing during incubation.