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Questions related to PCR Analysis
Referring to the attached image, has the RNA extracted from the FFPE sample been degraded? The lanes of agarose gel are as followed: 1) Lane 0: 50bp ladder 2) Lane 1: negative control (-RT) 3)...
02 February 2015 7,302 5 View
I'm a tech who's still getting the hang of running PCR. I've been doing it for about 6 months with three different genes (mop3fx, cre, and per2luciferase) and I've noticed that my mop3 primers...
01 January 2015 1,100 9 View
Hi, I'm trying to amplify a DNA fragment using primers with Tm: 79.3℃ and 63.4℃. I have tried T gradients between 54-68℃. There's always a loylt of unspecific bands. Ithink that a touchdown PCR...
01 January 2015 9,374 4 View
I want to do RT PCR of mRNAs of Heat Shock Proteins related to Drosophila tissue system. What is the procedure of sample preparation for RT PCR in insect tissue system?
01 January 2015 2,276 1 View
Hi, Could anyone explain how the coverage is calculated in amplicon seq? Since a target is PCR amplified and sequenced and there is a lot of duplication and no/less unique reads. Thank you, Ash
01 January 2015 3,940 2 View
Hello! I am trying to sequence (Miseq) a custom amplicon from many samples using the Nextera indexes. For those of you not familiar, this involves a two step PCR protocol. The first round of PCR...
01 January 2015 6,355 6 View
I want to quantify global DNA methylation in amphibians (toads) using MS-AFLP. I get PCR product in both the preselective and selective PCR, but I get no difference in the number of peaks/bands...
01 January 2015 6,112 13 View
i had run the std by using taqman probes by using same dilutions made from 100% methylated sample. but the pcr effiency is poor. so should i have to make the dilutions from same sample or from...
01 January 2015 7,978 3 View
What is the best PCR condition (PCR mix and temperature cycles) for amplification of the matK gene with matK-S51F and matK-1210R or matK-9R primers for Myrtaceae family (genus Metrosideros)? I...
01 January 2015 6,405 5 View
I want to have a good publication of analytical aspects of quantitative PCR not RT PCR. I need to know whats the dynamic range and how it can done best?
01 January 2015 422 5 View
what is the procedure and methods for that and what is there effectivness?
01 January 2015 2,637 1 View
I made a pcr reaction. I want to expect the size of the product recation. I have no reference except the guide paper
12 December 2014 10,425 22 View
I have a PCR product with 260bp size , usually i run it at 2% Agrose electrophoresis and it gave me a clear band as it appear in the lower line of the attached photo. After digestion , the...
12 December 2014 9,815 22 View
Two tissue sections are sent to lab for detection of mycobacteium tuberculosis in endometrium, histopathology report shows Granulomatous endometritis and PCR shows negative for mycobacterium...
12 December 2014 8,730 8 View
I used NaCl extraction of DNA in genus Gammarus of Amphipoda. Though my concentration and 260/280 readings are really good, my 260/230 is very low. Do I have contamination? If it's salt...
12 December 2014 2,914 2 View
I am trying to amplify seven SSR loci, that I used to genotype my study species individuals, in 2 out-group species (within same genus and within same family). I am getting amplification for 2...
12 December 2014 3,475 10 View
Usually, we perform SDS-Page in acrylamide gel and we use SDS buffer. If I use 1x TBE buffers for DNA or PCR products for running in acrylamide gel what will be the power/running condition?
12 December 2014 7,015 8 View
I optimized my PCR Reaction and it gave me very good result , after that some PCR product samples start to appear very faint in comparison with the other samples and the product of the first...
12 December 2014 9,524 14 View
Dear all, Good day and greetings. I'm just wondering if it's possible to perform PCR, once my cDNA fragment is inserted into Novagen's pET series vectors, using the above T7 pro/T7 term. as...
12 December 2014 5,788 4 View
Can anyone help in desigining primers for AGPase gene large subunit in wheat for real time pcr analysis ?
12 December 2014 9,559 3 View
Hi Guys, I've recently started doing ChIP-qPCR and have questions regarding analysis of data obtained from qPCR. My experimental design: I'm transfecting cells with a luciferase reporter plasmid...
12 December 2014 8,376 6 View
I follow Peterson et. al 2012's procedure for ddRAD-Seq. I ran PCR products-only and SPRI cleaned PCR products on a fragment analyzer, and seems like my PCR products are pretty clean and they are...
12 December 2014 8,724 7 View
I am trying to confirm the presence of a putative 8.5 kb insertion in a Drosophila mutant, by PCR and/or Southern Blotting. I use almost 10 ug DNA for each lane, and specific probes(812 bp) which...
12 December 2014 386 2 View
Hi researchers i plan to do a study of genetic polymorphism associating with primaquine therapy in our population. I was wondering how much of venous blood should i take from patients for DNA...
12 December 2014 7,433 5 View