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
1,991 views 3,271 posts
Questions related to PCR
I have cells with strong green and red/far-red autofluorescence. I need to culture them but I cannot find a good live/dead stain. Trypan Blue reacts with salts and proteins forming clumps. Red and...
05 November 2020 4,238 7 View
Strangely, my yeast colony PCR works only with Sapphire hot start master mix or Amplicon master mix but not by any other master mix, or if I individually add Taq polymerase with compatible buffers...
04 November 2020 3,984 3 View
Recently I've been doing some PCRs and the primes used to work a month ago, but now that we want to repeat the experiment, they are not working anymore. I have noticed something in my gels, and it...
29 October 2020 4,620 4 View
We are doing Coronavirus testing by real time PCR, targeting E gene (FAM), HKU (VIC) and Actin (ROX) gene. Real time PCR mater mix (20 ul) includes: Taqman Fast virus 1 step master mix (5ul),...
24 September 2020 5,975 3 View
I was doing the purification of Human Hsp40, and many papers refer to use detergent during the purification protocol. I just wanted to know how the detergent Triton-X or Brij-58 helps in avoiding...
11 September 2020 625 6 View
Pharmacogenetics
11 August 2020 7,443 1 View
N/A
09 July 2020 8,629 9 View
My labnet Multigene Mini Personal Thermal Cycler keep showing "room error" message when it was on. When I look at the troubleshoot manual, the manual state to reboot unit. However, I didn't know...
18 June 2020 1,769 3 View
I think the s.c. COVID 19 pandemic clinical manifestations (syndromas) have many different infective causes - Corona viruses (enteroviruses with many subtypes), othe viruses and bacterias! It is...
25 May 2020 5,978 5 View
Hello, Some references have mentioned the following equations for T/S ratio? The Relative telomere length from each sample, based on the T/S ratio, was calculated as the ∆Ct test...
11 May 2020 7,559 2 View
Dear colleagues working with RT-PCR devices, We are a team of data scientists, biologists, web developers working on an open-source machine-learning algorithm to contribute suppressing the COVID19...
07 May 2020 1,905 3 View
I used the Gibson assembly to join together two PCR fragments (4400 bp +770 bp). After transformation of E. coli with the Gibson mix, I isolated DNA from single colonies and I performed a...
29 March 2020 1,406 3 View
What is the time required for the body to produce a detectable concentration of anti SARS-CoV-2 ? Would such lag time make serological tests less interesting from a screening perspective? Should...
28 March 2020 9,467 3 View
I would like to analyze my qRT-PCR data generated from QuantStudio 5 PCR System using SYBR green method. I would like to import my raw data in readqpcr in R studio but I can not make data frame as...
27 March 2020 881 3 View
Traditional qPCR can give results in about an hour to detect/quantify DNA or RNA from various sources (bacteria, viruses, human, etc). I'm interested to know how you would use a machine that could...
25 March 2020 5,926 6 View
I have the following conditions: -perfect amplification -a 3000 bp amplicon -a %GC content of exactly 50% -15 cycles -0.5 mM dNTPs (A, G, C, T) -0.5 uM primers -10 ul reaction volumes I...
24 March 2020 5,085 3 View
Hello, I have a following issue: I extracted approx. 20 plant DNA samples from dry leaves (old herbarium items) using Qiagen DNeasy Plant Mini kit. When I run a PCR for housekeeping gene (alpha...
23 March 2020 2,772 6 View
I used BLAST to check for the specificity of my primers (for RT-qPCR) and could only find the desired product of 67 bp. After analysing with SYBR Green I foundnd two different melting curves for...
19 March 2020 5,169 4 View
My boss wants me to run PCR with a fluorescent tag. He says we can use the Tecan SPARK machine that we have for this. I am confused however. Do I run the PCR per usual with the thermocycler then...
16 March 2020 3,870 2 View
I know both can detect presence of virus but what is the exact difference between qPCR and qualitative PCR ? How and when is one preffered over the other?
16 March 2020 10,079 4 View
Hi I have a problem with optimizing TaqMan PCR? I am getting higher CT values in housekeeping gene GAPDH but undetermined target gene values. Also to be curious, I have ran those samples on 2%...
13 March 2020 7,574 7 View
Can anyone give the scientific explanation.
13 March 2020 7,453 2 View
I have been loading PCR product into 1.2% agarose gel and i've found that as the bands migrate down the gel (I let the gel run until the dye reaches the bottom edge) the bands become curved and...
13 March 2020 473 16 View
I have run a qPCR-assay using a Cy5-probe. On one of the plates, some technical replicates have very high RFU-levels, but the Ct-values of the replicates are almost identical. Does anybody know...
06 March 2020 6,662 4 View