How to select genes for qPCR validation in transcriptome/RNA seq data?

[@noorpk]

Thank you for uploading. It's useful.

[@asifmolbio]

Glad you like it

[@Yhusi]

Thank you for the knowledge. May I know how to calculate the expression level of certain gene from RANSeq data?I want to make ratio or certain gene compare to other certain gene to confirm my Western Blot data and my qPCR data. Thanks so much in advance for your help

[@asifmolbio]

How to calculate FC, log2FC, Pvalue, Padj, Up/down genes in RNA seq data using Excel
th-cam.com/video/HH3Mll4W5WE/w-d-xo.html

[@asifmolbio]

Please check this video hopefully will be helpful

[@MohammadSaleem-vl8sn]

Sir please make series of vedios starting from basics to advance bioinformatics/ biotechnology/genetics and both theory and practical examples and links for those software and materials which u describe

[@asifmolbio]

Thanks for suggestion. Here are already few uploaded th-cam.com/play/PL3fbGvDVm3usnd3Qidi5eX8azL-BUbZu2.html

[@ammarahchaudhary2820]

Aoa sir , plz made primer for RFLP and arms pcr in detail along with validation.

[@asifmolbio]

Wslam, sure i will upload. Stay tuned

[@poonam9511]

Hi doctor asif..I want to know how to create primer for gene in RNA seq data. Basically, I want to know to create primers in RNA seq data.

[@asifmolbio]

Poonam, usually you select gene’s ids from RNA seq data, not directly design primers from this data. I don’t know why you want to design primers from rna seq data

[@poonam9511]

@@asifmolbio i want to validate the up and down-regulated salient genes through PCR. how can i find primers for this data?

[@asifmolbio]

You can only select these genes based on RNA seq data. And design your primers by taking exon’s sequence/cds of selected genes using IgTDNA tool.

[@asifmolbio]

How to design primers for qPCR using online tool
th-cam.com/video/eKjv71j6UYI/w-d-xo.html

[@poonam9511]

@Dr. Asif's Mol. Biology hi sir, I have cds protein sequence . Could I use it for making primers by reverse translation and then use it for designing primer

[@pakchinalover]

How to make supplementary files?

[@asifmolbio]

See online version of any published paper in good journal like new phytologist

[@pakchinalover]

I am a newbie in this field, could you make a vedio on this? I didn't get what you have said.

[@dicenasi2194]

Hi Dr Asif, I'm new in NGS. My question is, how many genes are necessary to validate by qPCR? For publication. I saw you selected 5 for upregulation and 5 for downregulation to analyse by qPCR. Is there a minimum or maximum of genes to consider?

[@asifmolbio]

Hi, asi , There is no upper and lower limit but people usually take 5, 5. You can choose any other number of your choice.

[@snehaunni194]

Sir i am also working in plant transcriptome and ficused on a specefic serine/thr protein kinase. Sir pls help me to find out the isoforms of genes and how can i conduct a kegg pathway analysis.. And one more if possible pls give your whtspp number so that i can contact you. Actually i am writing a paper of my transcriotome work

[@asifmolbio]

Hi seha, please send the more details of your project to [e-mail]

[@snehaunni194]

@@asifmolbio i cant mail to this peculiar mail id. Thats the issue

[@asifmolbio]

Try this one [e-mail]