@reyamamlac

INTJ • DOHaD immunotoxicologist-wannabe but working on stem cell carcinogenesis/carcinogenic microenvironment, hematopoiesis, and extracellular vesicles

[@reyamamlac @reyamamlac]

@saee8778 @Jain_Aditi_ Agree. Funding and research facilities. I found out that these give more pressure than the advisor's pressure to do your research. PhD students have the tendency to go beyond what the advisor requires them to do so funding and research facilities matter a lot!

[@reyamamlac @reyamamlac]

@wu_zhaoming Can you share if you have access? I'm into prenatal inflammation exposure and disease development.

[@reyamamlac @reyamamlac]

@koo_lab Are PhD students eligible? I'm interested working with in vivo models related to stem cell carcinogenesis and stem cell niche.

[@reyamamlac @reyamamlac]

Known miRNAs in all samples is less than 500 out of 2656 species in miRBase. 3/n

[@reyamamlac @reyamamlac]

thread tweet #academictwitter #isev to small EV #sEV experts. My small #RNAseq data shows that I have around 4-5 million processed reads in the different samples. The mapped reads for known miRNA are 8-11,000 reads (0.17-0.25%). 1/n

[@reyamamlac @reyamamlac]

For prediction of known/novel miRNA, I get around 1.1-1.2 million mapped reads from the total 4-5 million reads. Unprocessed/untrimmed data shows 38-41 million reads. Do the numbers look good for differential miRNA expression analysis and downstream analyses? 2/n

[@reyamamlac @reyamamlac]

@GarciaLabMS I am working in a small lab and I have to do everything on my own. It's tiring and time-consuming to learn all the methods and analyze all the experiments on my own but I find it fulfilling when I learn from scratch and then perform successful experiments.

[@reyamamlac @reyamamlac]

@PhDVoice @Academic_Tasks @PostdocVoice Just raised her voice a little bit because I wanted to do something that the lab can't do. 😅

[@reyamamlac @reyamamlac]

@MachinePix She's hurting the robots!

[@reyamamlac @reyamamlac]

Today, I was offered a postdoc position by my PI but I don't know how to turn it down. I want to gain experience in a new lab in a different country where there is snow. I still have one year left before graduation and I'm worried she will take this negatively. #phdchat #phdlife

[@reyamamlac @reyamamlac]

@Iddomagen @BioRad @BioRadGenomics

[@reyamamlac @reyamamlac]

Any expert on #ddPCR for absolute quantification of #exosome #miRNAs? I got a series of questions since I'll be doing #ddPCR for the first time.

1. I understand that reference miRNA is not required. But I have 3 reference miRNAs available. Can I include them in my analysis?

[@reyamamlac @reyamamlac]

@Iddomagen Hi. I hope you can give comments/feedback for troubleshooting. I wonder why some of my targets give multiple bands in the positive events even though I did PCR optimizations. Some targets look okay but there is still rain. #ddPCR #extracellularvesicles #cellfreeRNA

[@reyamamlac @reyamamlac]

@SFUJessica @BinfoTrends Thanks!

[@reyamamlac @reyamamlac]

Is there a minimum number of small RNA sequencing replicates per experimental group for publication? Is duplicate acceptable? #smallRNA #exosomes #RNAseq #NGS #sequencing

[@reyamamlac @reyamamlac]

Details.

1. 10 ng starting sEV-RNA for RT-then diluted 10X
2. droplet prep-10 uL master mix + 2 uL universal primer + 2 uL LNA probe + 4 uL H20 + 2 uL cDNA =20 uL + 70 uL oil
3. 40 cycles PCR

[@reyamamlac @reyamamlac]

There's so much rain in my #ddPCR (first trial). Is this normal for ddPCR quantification of #smallEVRNA? Or do I need to do some adjustments? Any comments from the #ddPCR and/or #exosome experts?

[@reyamamlac @reyamamlac]

EV peepz are in Seattle while I'm still figuring out so many things in the lab about EV studies. 😅 I wish to attend to connect but...
#ISEV

[@reyamamlac @reyamamlac]

@AcademicChatter I like it when I think about the next set of experiments/conceptualizing future research. I don't like the feeling that I can never work on my ideas.

[@reyamamlac @reyamamlac]

@Iddomagen Thanks for the insights.

[@reyamamlac @reyamamlac]

@OGdukeneurosurg @NEJM Which countries have left driver's seats?