In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short... | Contact experts in PCR to get answers
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Questions related to PCR
I need to purify PCR product to further proceed for cloning, I dont have PCR purification kit so have to do manually. if I do Phenol:Chloroform:Isoamyl Alcohol 25:24:1 purification and ethanol...
09 November 2021 6,010 3 View
We are working with DNA samples extracted from faeces, so we are trying to find the optimal PCR conditions to amplify microsatellite markers, as the nature of the samples require quite an effort....
05 November 2021 6,839 9 View
I have just performed Direct PCR on some of my cyanobacterial cultures and would like to send the PCR product for sanger sequencing. I ran a gel and got very smeared bands. Do you think these will...
05 November 2021 1,808 5 View
Hi fellow researchers, I'm getting quite experienced now in 16S amplification and sequencing, however, I'm having strange results here. I've performed a large number of 16S PCR using the same...
02 November 2021 8,966 4 View
I'd like to concentrate my eluted plasmid DNA by heating it up in a thermocyler. The plasmid is eluted in water, and I don't have access to a speedvac right now. Would it damage my plasmid?
31 October 2021 4,589 6 View
I am think Multiplex is better in terms of optimization and cost but might not be better in terms of ease of doing/handling.
31 October 2021 6,863 4 View
I am doing over expression of an endogenous target gene in a CHO cell line. When I did PCR (whole length of the gene) to verify the existence of my target gene, I could only see the gel band when...
28 October 2021 6,447 6 View
Hi, I recently working on PCR-based site-directed mutagenesis, and I have some questions about the stability and storage of PCR products (before and after DpnI treatment). How long can I store...
23 October 2021 6,574 3 View
I need to diagnose HCV in infected patients by RT PCR but I am not sure of using either Serum or plasma as the sample. May commercial kits mention using plasma as the starting material but some...
22 October 2021 1,346 3 View
I need to incubate a large number of agar plates which contain bacteria for aerobic growth. If memory serves me right you should not have more than 8 plates in a single stack. Can someone please...
21 October 2021 3,413 4 View
The mean score was 93% on one test and 88% on the other.
19 October 2021 3,197 5 View
I work primarily with S. cerevisiae and commonly perform western blotting on total protein lysates. Our lab uses a fairly standard TCA/Bead lysis method...
17 October 2021 5,005 4 View
A DNA sample of 2000 base pairs was subjected to PCR under the given conditions Initiation; 950C for 5 minutes Denaturation: 950C for 45 seconds Annealing: 550C for 45 seconds Elongation: 720C for...
08 October 2021 3,184 5 View
i am facing a problem that previously in touch down and in gradient PCR of specific gene, specific band with non specific product come. but after 2-3 weeks when this same gene PCR was performed...
07 October 2021 7,080 3 View
In which concentration and volume (microliters) DMSO 5% or 1-10% I can used to optimize the PCR for specific product to avoid nonspecific product? I am facing a problem that I have use 1-10% and...
07 October 2021 9,017 3 View
Dear all, I want to do an EMSA assay using the LightShift® Chemiluminescent EMSA Kit. For that, I will add Biotin-target to the DNA by PCR using biotin-labeled forward primer. I would like to...
05 October 2021 4,264 2 View
A lab that works with ours does pretty frequent MiniSeq runs. I was going to sequence this 12 kb plasmid (which we have almost no information about) by Sanger sequencing, then ordering new primers...
01 October 2021 2,787 2 View
A 65 years old male patient, once a heavy smoker and HBV carrier suffers from some form of cancer with pericardial sac metastasis.The PET/CT shows there are four nodules with high SUVmax (higher...
30 September 2021 676 3 View
Hi everyone, I'm new in EMSA. And I found that Thermofisher offers a Chemiluminescent EMSA kit (catalog: 20158；https://www.thermofisher.com/order/catalog/product/20158#/20158). And I have few...
29 September 2021 3,593 2 View
my FW and REV primers Tm are 53.69 and 60.98 respectively based on the manufacturer's sheet( of course the TM(s) are different in NEB or primer blast). Unfortunately, my thermocycler does not...
26 September 2021 9,409 3 View
I isolated a number of entomopathogenic fungi on PDA in last three years, now I need to re-culture those organisms, but it fails, what method can I use to have them
25 September 2021 8,439 6 View
Hi, I am reading the article of BIOMED-2. There is something really confusing regarding the size of PCR product. Based on the design of the primer in TCRG tube B, the size range seems to be in...
24 September 2021 7,085 1 View
I have amplified ~800kb fragment and want to proceed with A-tailing of this product. How much template I need for this size. It did not work when I used 100ng of template which was a gel excised...
23 September 2021 3,759 1 View
I have a set of n aligned protein sequences. I have constructed a profile hidden Markov model (HMM) from the alignment with hmmbuild in the HMMER package. I would like to compare my constructed...
23 September 2021 7,098 5 View