טכנולוגיות מתקדמות

סיכום שיעור 9

Naser mondey

Naser mondey

פרטי הסיכום

קורס: טכנולוגיות מתקדמות

מספר השיעור: 9

סיכום השיעור

[11:51, 9.2.2021] Naser Mondey: – Not stable, degraded easily
– Only 3-5% of RNA is
mRNA. Most is rRNA
– Eukaryotic mRNA has
polyA tail at 3’
– There are also tRNA,
sRNAs, non-coding RNAs
-Nanodrop: spectrophotometer
260/230, 260/280 >1.8
Low 260/280 – presence of protein, phenol or
other contaminants that absorb strongly at or
near 280 nm.
Low 260/230- may indicate the presence of
contaminants which absorb at 230 nm such as
EDTA, carbohydrates and phenol.
[11:52, 9.2.2021] Naser Mondey: Only rRNA are seen (Why?),
indicating the RNA integrity
Cheap, but:
-Need to run ~500 ng
-Does not give an exact
value..
-Upper band should be stronger
-No Smear
-Not all kits include sRNA
-DNA( How do I know this is
DNA?)- Dnase treatment
[11:53, 9.2.2021] Naser Mondey: COMMON
APPLICATIONS
-Chromatin
immunoprecipitation on
Chip (ChIP on chip)
-Gene expression profiling
-SNP detection
-Alternative splicing
detection
DEEP SEQUENCING
Next Generation Sequencing (NGS)
High Throughput Sequencing (HTS
[11:54, 9.2.2021] Naser Mondey: MULTIPLE PLATFORMS
-Illumina (MiSeq, NextSeq, HiSeq, NovaSeq)
-Ion Torrent
-454
-Solid
-Nanopore/Oxford
-Pacific Biosciences (Long reads)
[11:54, 9.2.2021] Naser Mondey: ILLUMINA
APPLICATIONS
WGS- whole genome
sequencing
Exome sequencing
Target sequencing
ChIP sequencing
Methylation
sequencing
CRISPR
Single cell Seq
Bonus question-Exome and target
sequencing library prep is more
expensive so why do it?
Metagenomics Hint- Genome size Vs. Exome size
ETHICAL ISSUES
– $1000 human genome project:
Knowledge is power!
-Insurance
-Do you want to know if you will have a disease later in
life?

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