Biotechnology

• Tanu Singla

  Estimation of phenol content

  I have followed the folin ciocalteau method for estimation of phenol content using gallic acid as a standard but I could not find the way to interpret my results. I have made a standard curve andI have followed the folin ciocalteau method for estimation of phenol content using gallic acid as a standard but I could not find the way to interpret my results. I have made a standard curve and also taken the OD of samples but I do not know how should I proceed further. What are the calculations required for analyzing the results?

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  • 
    Tanu Singla replied

    Thanku 2 all for their valuable suggestions...:)

    8 minutes ago
• Juan Pablo Allocati

  I am designing a Biotechnology Lab from the ground up. Can you give me any advice?

  I have to design the different sectors, including the building. It's main activity will be plant genetic transformation. I would be glad to receive any publication or personal experiences.

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    Beyene Mekonen replied

    What I comment you is that before starting your activities, you should know what is biotechnology means and what the field needs to conduct the laboratory and the other is that what theWhat I comment you is that before starting your activities, you should know what is biotechnology means and what the field needs to conduct the laboratory and the other is that what the material/items and chemicals are needed to meet/ achieve your goal.

    3 hours ago
• Kaldeep Singh

  Restriction digestion of plasmid for gene extraction

  I have done restriction digestion many times but results are not become. How can I extract gene during restriction digestion of plasmid?

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    Marutpong Panya replied

    Please don't forget about the DNA methylation which will inhibit digestion. I have experience about DNA mehylation. My DNA could not be cut by restriction enzyme. So, I changed the bacterial host andPlease don't forget about the DNA methylation which will inhibit digestion. I have experience about DNA mehylation. My DNA could not be cut by restriction enzyme. So, I changed the bacterial host and finally the DNA/plasmid could be digested!!!!!

    5 hours ago
• Ibrahim Kutay Ozturk

  Could anyone with a lot of experience on real time PCR for relative expression (gene silencing) help me?

  I am working with Barley (Hordeum vulgare). I am trying to find out the silencing level of a gene via qRT-PCR. For that, I have completed the isolation of total RNAs for different samples, DNaseI am working with Barley (Hordeum vulgare). I am trying to find out the silencing level of a gene via qRT-PCR. For that, I have completed the isolation of total RNAs for different samples, DNase treatment and cDNA synthesis. I have also done qRT-PCR several times. My problem is this: Since the reference genes, which are housekeeping genes, have much more expression level than the gene of interest, I am obtaining Ct values at different regions. For example, I am obtaining Ct values of 25 when I use primers for reference genes, and 30 for gene of interest (GOI). I was informed that it is very important to obtain the Ct values in the same range. On the other hand, I have also read that using the same amount of cDNA at all wells is also essential. In my case, I can't do them both. I will try to change the normalization primers and look again, however, all the papers in my study area use 5-6 normalization primers. Thus, it is very likely that I will obtain similar results even if I change the normalization primers. Is it ok to use different amount of cDNA for different primer sets? For example, using 1 ng of cDNA when using reference primers, and using 4 ng of cDNA when using primers for GOI. If not, what do you suggest me to do? If you need any extra information, I would be glad to provide.

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    Stephen Doyle replied

    Check out the MIQE guidelines. Bio-rad have a nice little tech note on the topic http://www.sciencedirect.com/science/article/pii/S1046202310000204 Krzysztof is correct though - that is the wholeCheck out the MIQE guidelines. Bio-rad have a nice little tech note on the topic http://www.sciencedirect.com/science/article/pii/S1046202310000204 Krzysztof is correct though - that is the whole point of using reference genes. They should not be affected by the treatment, so they provide a stable reference between samples. Sometime finding a single reference that stays stable is difficult, so a few reference genes are used to normalise the data,

    15 hours ago
• Majid Shahbazi

  Could anyone let me know how to express a 32 amino acid peptide in E.Coli please?

  I want to produce and refold 32 mammalian amino acids that as a small protein in E.Coli. Has anyone got experience and suggestion which pET vector is suitable?

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    Majid Shahbazi replied

    Thanks all just would you all let me know do i need his tag or any specific tag? Tanks in advance for your kind consideration

    2 days ago
• Rima Jacob

  During DNA sonication, white fragments are appearing in the solution. Could that be DNA that is precipitating out?

  I am using 5ml DNA sample (dissolved in Tris-HCl ph=5.5). I need DNA of <200bp for which I sonicated it for 25mins (5secs on 10secs off, ampl 30% at 4C). I ran the sample on gel to optimize the timeI am using 5ml DNA sample (dissolved in Tris-HCl ph=5.5). I need DNA of <200bp for which I sonicated it for 25mins (5secs on 10secs off, ampl 30% at 4C). I ran the sample on gel to optimize the time of sonication. However after 15mins of sonication, fragments are formed in the sample which do not settle down with high speed centrifugation. Could it be DNA that is precipitating out? How should I troubleshoot?

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    Sakthikumar Ambady replied

    Rima, it seems like your sonicator probe is the sourse of this white powder. Clean the probe well or change it for a new one. At normal DNA concentrations, DNA should not be behaving this way. SinceRima, it seems like your sonicator probe is the sourse of this white powder. Clean the probe well or change it for a new one. At normal DNA concentrations, DNA should not be behaving this way. Since you see the white floating material in samples without DNA, it is definitely a sonicator problem.

    2 days ago
• Devi Prasad

  Ethidium Bromide degradation

  Most of us use Ethidium Bromide in labs. Are there any microbes that are known to degrade EtBr? If not till now, is it possible to produce such an organism?

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    Krishnan Nair replied

    EtBr is a potent mutagen and carcinogen. Handle it carefully. It can cross blood-brain barrier and so if ingested there is chance that it can even go to brain. All gels and solutions containingEtBr is a potent mutagen and carcinogen. Handle it carefully. It can cross blood-brain barrier and so if ingested there is chance that it can even go to brain. All gels and solutions containing EtBr should be treated with oxidizing agent to degrade it by oxidation. The best is to treat with KMnO4 not with hypochlorite. There were studies on the degradtion and efficacy of degradation products to act as mutagens/carcinogens in 80s. It was found that time that treating with KMnO4 is the best way to inactivate it. Now, regarding microbial degradation. I feel it is hard as EtBr intercalates DNA and interfere with its functions. The extend of inhibition of DNA functions depends on the amount bound per total genome DNA . This in a plasmid carrying strain, the plasmid replication is first interfered as the quantity of EtBr per genome is higher for plasmid than for the bacterial genome and that is why EtBr is used for curing plasmids. A bacteria in which the genome exists in multiple compartmens or partitioned in several replicons could be ideal candidate to look for as this will have higher resistance to the intercalating dye. Apart fron this, it should have genes encoding enzymes for the degradation of polycyclic aromatic hydrocarbons like in psuedomonas. May be one can introduce this by gene manupulations. Good luck in the study!.

    5 days ago
• Nandheesh P G

  Plastic Degradation

  Any known microbial groups or biological enzyme can degrade or reduce plastic ?

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    Andrea Zille replied

    Plastic is a very generic word. Without citing the well known biodegradable plastics (Polyhydroxyalkanoates, Polylcaprolactone, etc): - PET ca be partiallyPlastic is a very generic word. Without citing the well known biodegradable plastics (Polyhydroxyalkanoates, Polylcaprolactone, etc): - PET ca be partially degraded: http://www3.interscience.wiley.com/journal/108561288/abstract?CRETRY=1&SRETRY=0 - Nylon is well degraded by fungi: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=168325 - PVA is completely degraded by Pseudomonas sp. T. Suzuki, Y. Ichihara, M. Yamada and K. Tonomura , Some characteristics of Pseudomonas O-3 which utilizes polyvinyl alcohol. Agric Biol Chem 37 (1973), pp. 747–756. - Also aliphatic polyesters and polyurethenes are biodegraded: - Even one of the most stable polymers the polypropylene can be partially degraded : http://aem.asm.org/cgi/content/abstract/59/11/3695 Best Andrea

    11 days ago
• Abdul Mannan

  Is there any method to find the pI of a protein in solution by any chemical analysis in the lab?

  Actually i have purified my protein by considering pI from online calculation but i suppose to the pI of purified protein is different from online calculator. So that i want to check that..

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    Abdul Mannan replied

    thanks Rahul Raghvan., Mr. Tomáš Hluska and Birendra Singh for your good direction....

    11 days ago
• Fathima Sajina

  Can I work with Ethidium bromide on an open work bench without using a safety cabinet?

  Et Br is carcinogenic

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    Guido Vogel replied

    Avoid to use EtBr in the solid / pouder form. If possible always buy it as a solution (ready to use) so there is no need to weigh it in a safety cabinet. Inhaling EtBr as a pouder is the worst youAvoid to use EtBr in the solid / pouder form. If possible always buy it as a solution (ready to use) so there is no need to weigh it in a safety cabinet. Inhaling EtBr as a pouder is the worst you can do to your body. When handlich solutions containing EtBr allways wear gloves.

    13 days ago
• PANDIA RAJAN

  Preparing a slide for bacteria in SEM

  A bacterial broth containing bacteria should be checked for scanning electron microscope. How should the slide be prepared?

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    PANDIA RAJAN replied

    Thank u @ jayasankar

    13 days ago
• Mithat çelebi

  Which polymers can be used in biotechnological applications?

  Industrial biotechnology need hugely using immobilized enzymes. Can be used polymers for this area? Or other industrial biotechnology. In currently used, is there any industrial application ofIndustrial biotechnology need hugely using immobilized enzymes. Can be used polymers for this area? Or other industrial biotechnology. In currently used, is there any industrial application of polymers in biotechnology.

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    Mithat çelebi replied

    Dear Silva, Thanks for your help. Best Regards,

    14 days ago

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• Samani Tupufia

  How can we calculate the molecular weight of a yeast or protein?

  I am trying to calculate a molecular weight of a protien or yeast to see if Ican come up with a good mass balance when conducting reactions.

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    Lorena Pedraza replied

    Microorganism molecular weight is calculated by mean porcentual composition (C, H, O, N) and atomic weight of these elements. Average Yeast composition (C=47, N= 7.5%, H=6.5, O=31) from AtkinsonMicroorganism molecular weight is calculated by mean porcentual composition (C, H, O, N) and atomic weight of these elements. Average Yeast composition (C=47, N= 7.5%, H=6.5, O=31) from Atkinson and Mavituna, Biochemical Engineering and biotechnology Handbook.

    17 days ago
• Maniraj.R Mani

  I would like the sequence of one bacterial spe, anyone have an idea about the sequencing centers & cost in India?

  I would like to know about about microbial sequencing.

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    Chaitanya Joshi replied

    At Ome Research Facility, Dept of Animal Biotechnology, Anand Agriculture University, Anand, Gujarat we extend the facility on actual cost basis. We use two sequencing platforms -GS-FLX (Roche) andAt Ome Research Facility, Dept of Animal Biotechnology, Anand Agriculture University, Anand, Gujarat we extend the facility on actual cost basis. We use two sequencing platforms -GS-FLX (Roche) and PGM Ion Torrent (Invitrogen). Please refer J. Bacteriology for our three genome announcements

    18 days ago