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What are some hot biotech/biology research techniques being developed or used in labs
#1
At the DNA or RNA level, please. For example, the method of RNAi.

Thank you!
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#2
These are some of the recent breakthroughs or hot research going on in the Biotechnology sector that I stumbled across. There are lots more out there and many more to come.

1) Decay Fighting Microbes - Bacteria living in our teeth convert sugars to lactic acid, which erodes the enamel and eventually causes tooth decay. ONI Biopharm a Florida based company has come up with a new bacterial strain called SMaRT, that cannot produce lactic acid and it produces an antibiotic that kills the natural decay-causing strain. Dentists will only need to swab SMaRT, now in clinical trials, onto teeth once to keep them healthy for a lifetime.

2) Artificial Lymph Nodes - The Riken Institute scientists have come up with artificial lymph nodes, organs that produce immune cells for fighting infections. The artificial lymph nodes can be used as immune boosters. Doctors could fill the nodes with cells specifically geared to treat certain conditions, such as cancer or HIV.

3) Asthma Sensor - Asthma can be really fatal and accounts for most of the emergency visits in the US and worldwide, but scientists hope this situation will change with the new asthma sensor developed at the University of Pittsburgh. Inside the handheld device, a polymer-coated carbon nanotube—100,000 times thinner than a human hair—analyzes breath for minute amounts of nitric oxide, a gas that lungs produce prior to asthma attacks.

4) Cancer Spit test - Say bye to biopsies, University of California - Los Angeles have come up with a test which can detect oral cancer from a drop of saliva. Proteins that are associated with cancer cells react with dyes on the sensor, emitting fluorescent light that can be detected with a microscope. According to engineer Chih-Ming Ho the same principle could be applied to make saliva-based diagnostic tests for many diseases.

5) Biological Peacemaker - The good old fashioned electronic pacemakers are effective but they eventually wear out. The scientists have come up with a solution to combat this problem. Researchers at several universities are developing a batteryless alternative: pacemaker genes expressed in stem cells that are injected into damaged regions of the heart. Better suited for physical exertion, biological pacemakers have been shown to bring slow canine hearts back up to speed without complications.

6) Absorbable Heart Stent - The function of a stent is to open arteries that have become narrow or blocked as a result of a coronary heart disease. Drug-eluting stents release medication that keeps the artery from narrowing again. The bio-absorbable version made by Abbott Laboratories in Illinois goes one step further: Unlike metal stents, it does its job and disappears. After six months the stent begins to dissolve, and after two years it's completely gone, leaving behind a healthy artery.

7) Gastrointestinal Liner - Obesity is associated with type II diabetes, which over time wears out the pancreas. A gastrointestinal liner developed by Massachusetts-based GI Dynamics may restore the obese to a healthy weight by preventing food from contacting the intestinal wall. The Endobarrier is routed endoscopically through the mouth—unlike a gastric bypass, no surgery is necessary—and lines the first 2 ft of the small intestine, where the most calories are absorbed (nutrients are still absorbed farther down the intestine).

These according to me are by far the interesting and innovative research going on in the Biotechnology sector.
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#3
DNA/RNA associated methods and their application

Polymerase chain reaction (PCR)

PCR is one of the oldest techniques (developed in 1983.) used for multiplying DNA sequences. DNA piece could be amplified thousand to million times. It’s used for identification of hereditary disorders, for paternity testing or DNA profiling (in forensics), for DNA -based phylogeny and for diagnostics of infectious diseases. DNA piece that needs to be multiplied is mixed with heat stable DNA polymerase (usually Taq polymerase), DNA primers and nucleotides in the buffer solution containing certain amount of Mg2+ (or Mn2+) and potassium ions. Initial step is DNA melting where high temperature is breaking double DNA strand in two pieces. After separation, DNA primers are attaching to the single stranded DNA and assembly could start. Each DNA piece serve as template for DNA polymerase that is adding complementary nucleotides from the solutions until new double helix is created. By rising the temperature, cycle can start all over again. It is usually repeated for 20-40 times and in each cycle amount of DNA product is doubled.

Quantitative PCR (qPCR) or real-time polymerase chain reaction

This method is simultaneously amplifying and quantifying produced DNA. Oligonucleotids in solution are labeled with fluorescent dyes that could be visualized after hybridizing with the complementary nucleotides on the DNA. This method is mostly used in research and for diagnostic purposes (for cancer, infectious diseases, flu…).

Reverse transcription polymerase chain reaction (RT-PCR)

In RT-PCR method, RNA molecule is reversely transcribed into its DNA complement (cDNA) using enzyme reverse transcriptase. cDNA is further amplified using PCR. This technique is common when viruses are under investigation, as most of their genomes are consisted of RNA.

Gel electrophoresis

Gel electrophoresis is used for separation of the molecules based on their size and electric charge. Porous gel is usually made of agarose or polyacrylamide. Potato starch could also be applied. When gel is placed in the electric field, movement of the negatively charged nucleic acids will depend on their size. Shorter pieces will move faster and pass greater distances on the gel compared to long pieces, allowing quick and efficient separation of the mixed population (RNA and DNA) of the nucleic acids. Visualization of the separated molecules is achieved using ethidium bromide for DNA and RNA (provides blue fluorescence under UV light), or silver stain or Coomassie Brilliant Blue dye for proteins. Main applications of gel electrophoresis are in PCR products analysis, in restriction mapping of cloned DNA, for the separation of the restricted DNA pieces prior some other laboratory analysis…

Macromolecule blotting and probing

Southern blot method is used to detect specific DNA sequence in the sample. It combines gel electrophoresis (for DNA pieces separation) and probe hybridization (to determine if sample contains DNA complementary to the probe). Main applications are in homology based cloning, for identification of the methylated sites in certain genes or in knockout stem cell engineering.

Northern blotting is used to detect RNA in the sample. It provides information on gene expression patterns in different tissue/organs, in various developmental stages, during the course of the treatment….

Western blotting, known as protein immunoblot, is used to detect protein in the sample. Specific antibodies are used for targeted proteins after they are separated on the gel. This method is applied for HIV, Bovine spongiform encephalopathy, Lyme disease or Hepatitis B infection testing.

DNA microarray (DNA chip, biochip)

This technique is used when multiple genes expression need to be measured simultaneously. Each gene spot contains specific DNA sequence (probes) that are used to hybridize cDNA or RNA (target) under specific conditions. Probe-target hybridization is detected by labeling the target with fluorescent dye. Array can contain thousands of genes and parallel experiments could be easily performed. This method can be used for genetic profiling, comparative genomic hybridization, single nucleotide polymorphism detection, selection of the appropriate drug candidate….

Genome mining

New technique in extracting genomic data resulted as collaboration between scientist from the Scripps Institution of Oceanography and The Skaggs School of Pharmacy. Raw biological material could be evaluated for the known and unknown peptides using novel mass spectrophotometer. After peptides are fragmented to their amino acid building blocks, scientists could map them to the genome level. This method is unique, as it allows tracking “biosynthetic pathways”, or how molecules are reassembled (all the way from genomic expression to the end-product development). Two new classes of peptides are discovered using this technique. Information on “biosynthetic pathways” is valuable in a lot of fields, especially in pharmaceutical industry.
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What are some hot biotech/biology research techniques being developed or used in labs00