11 Questions 24 Answers 0 Followers
Questions related from Emma Monaghan
I was just wondering what your thoughts were on storage of purified PCR products - fridge or freezer temperature? I always elute purified products using the buffer provided with the purification...
04 April 2014 7,699 5 View
I'm a bit confused by my cloning PCR results. I have used the TOPO TA Cloning kit and everything seemed to go to plan - I got colonies, I grew them in the indicated liquid culture and then...
04 April 2014 3,476 8 View
I have been using a standard hot-start Taq polymerase for processing samples for NGS. My samples are low yield so when working with them before NGS preparation, this mix with the standard...
04 April 2014 3,001 2 View
I have some liquid cultures of individual colonies that have been grown in LB plus kanamycin [50ug/ml]. I want to extract plasmid DNA from them all in one big batch. To do this, I was going to...
03 March 2014 2,285 4 View
I have read that repeated freeze-thawing can damage DNA but then a lot of kit guides recommend freezing for storage. I have stored PCR products and purified PCR products in the fridge and found...
11 November 2013 9,816 14 View
Hi everyone, I am currently looking at trying to improve a DNA extraction method we use within my lab group. In the lysis stage of the extraction, we add phenol, 20% SDS and sodium phosphate pH 8...
11 November 2013 8,482 7 View
I am currently developing a protocol to process approximately 400 samples using the Illumina MiSeq platform. We are using the two-step PCR approach and have designed our own primers for this....
09 September 2013 3,832 3 View
I have ran a 16S general bacterial PCR at both 30 and 35 cycles as I have read on ResearchGate previously that people find 30 cycles achieves good amplification with no non-specific...
08 August 2013 6,777 22 View
Hi all, I am completing a 16S general bacterial PCR where I am using 1ul of 1:10 diluted PCR product as the DNA template for each sample I am processing. I have ran the PCR at 35, then 30 and now...
08 August 2013 5,575 29 View
I am determining my protocol to run a denaturing DNA PAGE gel to look at DNA fragments just under 100bp. I was wondering whether people commonly used a stacking gel to enable a good transition of...
04 April 2013 4,056 3 View
I am analysing data from 30 DGGE images using GelCompar. The banding patterns are quite complex so I have been doing a curve-based analysis. The methodology in published literature seems to be to...
01 January 2013 9,853 3 View
