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1 What direction do you think is of greater potential in biofuel-related studies?
By Daniel Deng · University of Southern Mississippi

Tarun Kant · Arid Forest Research Institute, India

I fully agree with you observations and conern Daniel. I feel, a lot of work has been done on different ways of biofuel production. But emphasis on bi... [more]

I fully agree with you observations and conern Daniel. I feel, a lot of work has been done on different ways of biofuel production. But emphasis on biofuel refinement and release of a final good quality, readily acceptable biofuel needs more research. Not much work on blending of biofuels from various sources has been done so far. A good blend may iron-out shortcomings of individual biofuels from different sources. This might be an area that may be explored. Work on large scale production and refinement is yet to be tested in a real world business model. Its time, people get biofuels on a sustainable basis in gas stations instead of just in magazines and journals.

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5 What amount of cells at a density of 2.0x10power5 is suitable for RNA harvest
By Stephen Karori · Egerton University

Muhammad Imran · Ajou University

Trypsinization followed by deactivation and washing the culture dish with the same media will perfectly work well. You can easily check the adherent c... [more]

Trypsinization followed by deactivation and washing the culture dish with the same media will perfectly work well. You can easily check the adherent cells (if any) through microscope. Centrifugation followed by washing with PBS is highly recommended. I would suggest you to dissolve th pellet in Trizol and process it directly or store it at -70C for later use rather than storing the pellet. For RNA preparation you can harvest your cells directly with trizol as Sagar rightly explained. To this end you remove the culture media and wash the cells with 1X PBS (2-3 times) followed by the addition of Trizol directly to the culture dish. Kepp it at room temeperature for short time (along with gentle rocking or pipetting to dislodge and dissolve the cells) and then process it as described previously. 1x10^6 cells per sample are suffcinet enough to give you a good amount of RNA pellet. For cells to Trizil ratio please check your kit instructions. Good luck.

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200 Concentration of glycerol used for storage of bacteria?
By Sophia Wang · Xiamen University

Vincent Chiang · National Defense Medical Center

In our lab, we use 50% of glycerol (400 ul) and mix with bacteria (600 ul) to make the final v/v of 20% then freeze in -80 degree.

In our lab, we use 50% of glycerol (400 ul) and mix with bacteria (600 ul) to make the final v/v of 20% then freeze in -80 degree.

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Question:
New Scale up Process
i am doing project in waste water treatment using bacteria in 5L stirred tank reactor. i have to Scale up this process......... what are the parameter... [more]

i am doing project in waste water treatment using bacteria in 5L stirred tank reactor. i have to Scale up this process......... what are the parameters have to consider for scale up and how to do it?

By Puhazh Selvan · Anna University of Technology, Coimbatore

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New What are the GMO-related legal aspects in strain engineering for microorganisms used for food products?
I've been told that, for example, people use genetic engineering to generate strains that have the desired properties, analyze what changed, and then ... [more]

I've been told that, for example, people use genetic engineering to generate strains that have the desired properties, analyze what changed, and then use methods that won't necessitate a GMO label to try to reproduce those changes in a strain that will actually be used to make food/beverages. I would really like to find a reference that discusses this, but I'm having trouble finding one. Thanks!

By Peter Enyeart · University of Texas at Austin

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146 Are you ready to eat a 100% tested safe GM food crop?
By Vageeshbabu Hanur · Indian Institute of Horticultural Research

Max Chartrand · Northcentral University

I agree, DR. Maize is pretty benign in the essential and micro nutrient department. On our end we see the effects of various diets at the practice lev... [more]

I agree, DR. Maize is pretty benign in the essential and micro nutrient department. On our end we see the effects of various diets at the practice level and try to document those differences in qualitative terms that can be someday allow us to design a quantitative format. I believe the recent studies on GMO high frustose are pretty conclusive that health effects do occur that should serve a cautionary note to marketers of mass produced food--we stop the HFCS and people lose weight and blood sugar spikes stop. All the way around, conducting even metastudies from the existing literature seem almost impossible. My contention all along is that independence in studies is a vital component of objectivity--hence, the need for blinded funding and researchers who can get past the more sterile view of nutrition ("it looks like, acts like, and therefore, must be"... vs "what does the body say about it"--proinflammatory vs anti-inflammatory cytokines). We see a truly organic diet (without aggressive cooking and microwaving) to produce anti-inflammatory responses versus the inflammatory responses typically seen in GMO diets. But to be convincing to the entire scientific community, and eventually the consumer public, more studies are needed looking at this in strictly objective terms.

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3 What are the current main problems in processing lignocellulosic biomass to 2nd generation fuel?
By Anna Lekanova · Kajaani University of Applied Sciences

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New In using the haldane equation to detemine inhibition, how do i detemine Ki and Ks in microbial kinetics
In using the haldane equation to detemine inhibition, how do i detemine Ki and Ks in microbial kinetics

In using the haldane equation to detemine inhibition, how do i detemine Ki and Ks in microbial kinetics

By Obinna Oje · University of Nigeria

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2 I want to know what is +1 value and -1 values for response surface methodolgy software?
By Margi Shelat · Kadi Sarvi Vishwavidyalaya University

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1 Can anyone help me to find the best protocol for hairy roots induction using agrobacterium rhizogenes?
By Febri Vidiyanti · Airlangga University

Annick Keyser · Ghent University

We use a modified protocol of Boisson-Dernier et al. 2001 What did we modify? - we use YEB medium instead of TY medium for the A.rhizogenes. We put a... [more]

We use a modified protocol of Boisson-Dernier et al. 2001 What did we modify? - we use YEB medium instead of TY medium for the A.rhizogenes. We put a 2ml ON culture on a YEB-plate to grow a layer of A.rhizogenes containing our construct. - we use Soli medium supplemented with 1mM NH4NO3 instead of Fahraeus - we do not use Km in the plates where we put the cut/infected seedlings. Our constructs have a gfp-cassette so we can cut away non-transformed roots under a gfp-binocular - we seal our square plates with Micropore tape instead of Parafilm so we don't have to make incisions in the tape for gas-exchange - after 5 days in the 20C room for the co-cultivation we transfer the seedlings to new Soli+ plates and from then on we grow the roots between 2 layers of Anchor germinaton paper @24C, 16h photoperiod. This paper makes it more easy to cut away non-transformed roots

http://www.molecularinfo.com/MTM/K/K3/.../K3-1-7.pdf‎ ×

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109 Is the relation between OD reading and cell concentration (cells/mL) of bacteria different in various culture media?
By Hamed Khodadadi · Eastern Mediterranean University

saravanan.c Kalki · Pondicherry University

Dear Sir, kindly measure the any uninoculated broth in spectrophotometry because that turbidity involve in the growth OD the cell and minus it with ur... [more]

Dear Sir, kindly measure the any uninoculated broth in spectrophotometry because that turbidity involve in the growth OD the cell and minus it with ur results.same time do CFU,PCV,DBM for urs desirable organism which support the conc of the cells.

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2 Can anybody explain poultry litter for biogas production tips to follow smooth operation
By Sataluri Ayyangar · Nava Bharat Ventures Limeted

Vijayaraghavan Gonuguntla

CONSORTIUM of Acetobacter xylinum Desulfovibrio desulfuricans One of the following Methanomicrobium mobile. Methanobacterium formicicum Methanobac... [more]

CONSORTIUM of Acetobacter xylinum Desulfovibrio desulfuricans One of the following Methanomicrobium mobile. Methanobacterium formicicum Methanobacterium thermoautotrophicum Marburg Methanobrevibacter ruminantium, Methanococcus voltae Alongwith enzymes like Amylase, Protease, Lipase may be used

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5 Is there any easier method other than silver staining for staining DNA fragments in polyacrilamide gel?
By Saba Saboori · University of Tehran

Marko Dolinar · University of Ljubljana

The cheapest and easiest way is by using ethidium bromide. Let the gel stain for a few minutes in a standard EtdBr staining solution (just as for agar... [more]

The cheapest and easiest way is by using ethidium bromide. Let the gel stain for a few minutes in a standard EtdBr staining solution (just as for agarose gels) on an orbital shaker. Wash with tap water briefly. Be careful not to tear the gel. If you don't see bands, continue staining for another 5-10 min.

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Question:
New Please can anyone give me details of how to produce industrial Enzyme(from business perspective, how to desgin, how to run, cost analysis)
I want to do business specially in enzyme producing industry, so please help me by giving some suggestions or give me a opportunity to visit any enzym... [more]

I want to do business specially in enzyme producing industry, so please help me by giving some suggestions or give me a opportunity to visit any enzyme industry in India.

By Utpol Das · South Asian University

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2 Why does DNS precipitate when added with sodium acetate buffer pH 4.8, while doing enzyme assay for finding glucose concentration?
By Nisha Venkatesh · Anna University, Chennai

Daniel Deng · University of Southern Mississippi

For reducing sugar assay, the DNS solution contains NaOH which makes the reaction buffer highly alkaline.

For reducing sugar assay, the DNS solution contains NaOH which makes the reaction buffer highly alkaline.

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6 Can one ferment a particular substrate with a combination of yeasts and bacterial cells considering the differences in their growth requirements?
By Adebare Adeleke · Modibbo Adama University of Technology, Adama

Muhannad Massadeh · Hashemite University

I suggest to use a multistage system to give better chances for each microorganism to grow

I suggest to use a multistage system to give better chances for each microorganism to grow

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3 EDC chemical handling
By Shazzy Drogba · Universiti Malaysia Pahang

Bilal Demir · Ege University

EDC/NHS couple needs fresh medium. You cannot rely on stock solutions especially these solutions (but not buffers ofcourse and cell culture mediums)

EDC/NHS couple needs fresh medium. You cannot rely on stock solutions especially these solutions (but not buffers ofcourse and cell culture mediums)

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18 How to prepare a 0,2 M phosphate buffer (Na2HPO4-NaH2PO4), pH 6.4?
By Valeria Scagliotti · Università degli Studi di Milano-Bicocca

Trung Kien Nguyen · University of Incheon

You can make sodium phosphate buffer directly from purity sodium phosphate powder by go to sigma website and search for sodium phosphate buffer. Good ... [more]

You can make sodium phosphate buffer directly from purity sodium phosphate powder by go to sigma website and search for sodium phosphate buffer. Good luck

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1 Why frequency shift for Neutravidin in QCM-D is not stable? how we can make Neutravidin solutions?
By Behnoosh Tajik · University of Melbourne

Jeffrey Brender · University of Michigan

You likely have additional layers of neutravidin bound on top of the first layer. Try a wash step to remove non-specifically bound avidin as recommend... [more]

You likely have additional layers of neutravidin bound on top of the first layer. Try a wash step to remove non-specifically bound avidin as recommended in this paper. http://www4.ncsu.edu/~ojrojas/PDF/2012_22.pdf

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1 Does anybody have whole sequence information of pART7 vector?
By Hana Zandkarimi · University of Tehran

Sadeeq ur Rahman · VU University Amsterdam

http://nanolight.com/Uploads/121%20%20pART27%20info%20PDF.pdf. this is the general map that would help you out for cloning but if you need the whole s... [more]

http://nanolight.com/Uploads/121%20%20pART27%20info%20PDF.pdf. this is the general map that would help you out for cloning but if you need the whole sequence you need to ask it from the provider

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7 DNA eluted with 6M Guanidine hydrochloride from Streptavidin Magnetic beads, does it need further purification for downstream applications?
By Sudip Das · Universität Würzburg

Francesco Reddavide · Technische Universität Dresden

Hello Sudip, I see that the Roche Streptavidin beads have a capacity of 1800 pmol/mg for free-biotin. I suggest to load three times, or more, this am... [more]

Hello Sudip, I see that the Roche Streptavidin beads have a capacity of 1800 pmol/mg for free-biotin. I suggest to load three times, or more, this amount (it depends how much beads you used) then you can take the supernatant. I don't think the free biotin can inhibit PCR, but you can also remove it. Alternatively, you can try the invitrogen protocols for the Elution. Best, Francesco

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36 Storage of primers
By A.K.M. Moyeenul Huq · National University of Malaysia

Shiau Foon Tee · Universiti Tunku Abdul Rahman

I agree with researchers to dilute primers in TE buffer and keep in -20 (100uM )for a long term use (stock solution). I aliquot it into several tube... [more]

I agree with researchers to dilute primers in TE buffer and keep in -20 (100uM )for a long term use (stock solution). I aliquot it into several tubes for preparation PCR. The product of PCR that I got was in good quality. This showed that primers work well... Hope this will help u

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2 Conjugative transfer of Plasmid (pAKE604) from Ecoli S17 to Rhodococcus (Gram+), I need your recommendation..?
By slimane khayi · Université Moulay Ismail

Przemyslaw Nuc · Uniwersytet im. Adama Mickiewicza w Poznaniu

If You want to transfer a plasmid, try this: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC332450/

If You want to transfer a plasmid, try this: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC332450/

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7 Buffers for biology
By Ahmad Ahmad Sobri · University of Bristol

Ananda Ayyappan J.V · Heinrich-Heine-Universität Düsseldorf

the answer is NO! the buffering activity of any buffer depends on its pKa as said above. if you are trying to make an acidic pH with a buffer that has... [more]

the answer is NO! the buffering activity of any buffer depends on its pKa as said above. if you are trying to make an acidic pH with a buffer that has basic pKa you will end up making buffer with false pH.

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2 Preventing Repeated Strain Injury (RSI ) other than ergonomics, caused due to working for long hours with micro pipettes . Is it possible?
By Abhinay Batchu · Indian Institute of Chemical Technology

Abhinay Batchu · Indian Institute of Chemical Technology

thanks to Enrique, i came across this very good journal, Applied Ergonomics by SD http://www.sciencedirect.com/science/journal/00036870

thanks to Enrique, i came across this very good journal, Applied Ergonomics by SD http://www.sciencedirect.com/science/journal/00036870

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4 Cell-free or bacterial expression? Which of the methods is advantageous in obtaining a recombinant disulphide-rich protein?
By Peter Dubovskii · M.M. Shemyakin–Yu.A. Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences

Peter Dubovskii · M.M. Shemyakin–Yu.A. Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences

Thank you!

Thank you!

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16 How to preserve the cut band from 1% Agarose gel before purification?
By Pardis Fazli · Putra University, Malaysia

Pardis Fazli · Putra University, Malaysia

Thank you very much for answering, how long may the bacterial DNA samples be stored as gel fragment in -20oC?

Thank you very much for answering, how long may the bacterial DNA samples be stored as gel fragment in -20oC?

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15 Apart from microbiological and biochemical tests, how can bacteria be characterized by molecular methods?
By Aparna Sekhar · Indian Institute of Horticultural Research

Ana Henriques · Universidade do Minho

Molecular techniques that could be useful could be: DGGE, RFPL, PCR, Sequencing coupled with comparisons of results of previous studies can give you s... [more]

Molecular techniques that could be useful could be: DGGE, RFPL, PCR, Sequencing coupled with comparisons of results of previous studies can give you some more information about the characteristics of your microorganism.

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8 Why do you need to centrifuge the master mix before pipeting it to individual pcr tubes?
By Carlo Antipuesto · Ateneo de Manila University

Vasudevan Rajeswari · Madurai Kamaraj University

In order to pool all the contents to the bottom of the pcr tube or eppendorf tube. This would certainly avoid any of the pcr components sticking to an... [more]

In order to pool all the contents to the bottom of the pcr tube or eppendorf tube. This would certainly avoid any of the pcr components sticking to any one corner of the tube. This would also ensure proper mixing of all components. But make sure that you go for a quick centrifuge to avoid deactivation of taq polymerase enzyme.

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6 I need info on Balsamic Vinegar analyses. Can one use the same methods as we use for wine?
By Valmary van Breda · Agricultural Research Council, South Africa

Maria Gullo · Università degli Studi di Modena e Reggio Emilia

you need also to determine ethanol consumption. Enzymatic assays or distillation can be useful.

you need also to determine ethanol consumption. Enzymatic assays or distillation can be useful.

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