Jawwad Ahmad

9 Questions 137 Answers 0 Followers

Questions related from Jawwad Ahmad

What impact would be there on RNA-SEQ experiments if we do not use any RNA Spike-In controls (like ERCC or SIRVs)?

Hi everyone, if I do not use spike-in RNA isoform controls for a customized RNA sequencing experiment (not the whole transcriptome, just based on few specific genes we are interesting), so is...

02 February 2020 8,155 2 View

Can I add "enzyme-based RNAse Inhibitors" to the extracted FFPE RNA elutions to avoid any possible RNA degradation by RNAses?

Hi I am using RNAse Inhibitors (applied biosystems), which are enzyme-based and are meant to add during cDNA synthesis phase as per its manufacturer policy (1ul to each 20 ul RT reaction). But,...

05 May 2019 9,091 2 View

Is there any undesirable effect of bacterial RNA load of normal intestinal flora present on human colon samples for the gene expression analyses?

It is a known fact now that interactions between bacterial (normal flora) and colon cancer cells influence the transcription profile of host cells or enterocytes. So, how can I avoid or minimize...

10 October 2018 6,170 1 View

Can we pool together the same type of samples in qPCR for delta-delta ct method?

Is it worthy to pool all the samples together belong to a same group, but originated from different animals of a same species, to save time and resources (reagents)? For example; From my negative...

06 June 2018 8,663 7 View

Does "No Primer Dimer Formation" guarantee the absolute utilization of entire primer content (F & R both) in a PCR reaction?

Dear all, in a PCR (conventional & realtime both), if we achieve our desired product (GOI) and along with, there is also no primer dimer (PD) formation seen on the gel and melt curve, does...

11 November 2017 4,512 4 View

How Spermidine alleviates in vitro full-length cDNA synthesis from RNA?

Spermidine through which mechanism, behavior or pathway results in stable full-length cDNA from the RNA when is used with certain reverse transcriptases, like AMV in RT step. What is its actual...

11 November 2017 6,067 2 View

How to get rid of PCR amplification artifacts while using the Taq DNA polymerase in PCR to prepare the libraries for NGS applications?

Taq DP is very well known to induce Indels and single base substitutions etc. So if any polymorphism or genetic variation detected later on, through any NGS platform, what makes us sure that these...

05 May 2017 4,200 4 View

How an Asymmetric qPCR (unequal forward and reverse primers) results in an improved Melt Curve?

A low concentration of a primer than the other one in high concentration or even in volume often exhibits good melt curve appearances. What could be the reasons behind this? Is this dependent on...

11 November 2016 5,040 3 View

The frequency of distribution of CRISPR elements among Gram -ve and Gram +ve bacteria almost the same or more in G +ve than G -ve or vice versa?

Also what about CRISPR-Cas9 presence level in Atypical bacterial genera?

11 November 2016 4,358 0 View