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Questions related to Molecular Biological Techniques
I have isolated RNA with high purity and concentration from banana and converted it to cDNA then amplified with reference gene (Actin, tubulin, ubiquitin) and specific primer (no cross dimer, hair...
28 July 2016 1,415 3 View
Hi, I'm referring to a paper 'Studies on the “Insoluble” Glycoprotein Complex from Human Colon IDENTIFICATION OF REDUCTION-INSENSITIVE MUC2 OLIGOMERS AND C-TERMINAL CLEAVAGE' from Herrmann et al...
28 July 2016 7,771 2 View
I did a RNA isolation with a Polysome Pulldown + TriZol, followed by a purification with a RNeasy Kit. With the isolated RNA a transcriptome should be generated. My samples on the Nanodrop looked...
27 July 2016 5,149 3 View
I am doing a PCR of my vector. I expect the PCR product to be of 5200bp. The PCR of the vector works only with 2ug DNA and Phusion Polymerase. It does not work at all with lower concentration of...
27 July 2016 5,068 3 View
Hi, I am considering to buy MAX Efficiency DH5 alpha Competent Cells to try to do both electroporation and heat-shock experiments with large plasmids. I don't know what procedure will be better,...
27 July 2016 9,903 6 View
Hi all, So I want to clone out a gene from a commercially obtained cDNA library. I was wondering if people have been successful using a nested primer approach for a gene of almost 6Kb? I have...
27 July 2016 1,816 3 View
I have an insert and vector with one sticky end and other blunt end, It did work when insert size was small but at the time of large size fragment it didn't. Can anybody provide me solution?
26 July 2016 2,563 5 View
A typical digestion requires 1ug DNA for a 50uL reaction which results in DNA that is 20ng/uL. Once I do a cleanup the concentration is too dilute for NGS (or
26 July 2016 9,121 5 View
Any suggestions are more than welcome.
26 July 2016 8,281 3 View
Datasheet (Nanodrop 2000) and others said possibly these peaks are due to phenol contamination. Methods like 1- Butonol/ sodium acetate re-precipitations have shown to improved the purity of RNA....
26 July 2016 8,985 14 View
I want to add Ferrozine to my YPD agar medium. I wonder whether Ferrozine is stable with heat (autoclave) and light. If you used to use this kind of medium, please teach me! Thank you so much!
26 July 2016 8,128 2 View
In studying of bacteria molecular characterization and antibiotic resistance, It is important to do the PCR for the bacterial genes and resistance genes before sequencing or Direct sequencing from...
26 July 2016 5,030 2 View
I want to ligate an insert(1270bp) into pet32a .pet and insert digested with NcoI and XhoI.
23 July 2016 1,521 3 View
My protein shows an increase in tyrosine fluorescence prior denaturation transition in a titration with GndHCl. But it decreases it´s fluorescence prior the denaturation transition in a titration...
23 July 2016 5,208 2 View
Hello everyone, I have some problems about designing functional genes specific Taqman qPCR primers/probe. Any suggestion is helpful! I'm planning to design a set of Taqman qPCR primers/probe to...
22 July 2016 4,509 3 View
i need to know the embroy satge at which the drugh should to induced.
22 July 2016 920 3 View
Factors affecting somatic cell count in milk.
22 July 2016 8,014 3 View
I need a viral lysate. There are some substance to it? Something that disrupts the lipid envelope.
21 July 2016 4,822 1 View
I have been trying to detect mineralisation in human osteoblasts and the hFOB cell line for some time now, but have been unsuccessful. My old method was to seed out 20,000 cells/cm2 and leave them...
21 July 2016 8,959 2 View
My POI is a human protein. However we when we had the DNA sequence synthesized we codon optimised for S.cerevisae (for protein expression). However, I now wish to do some interactome studies in a...
21 July 2016 1,547 3 View
I need remove cell debris, however i don't know if can filter the dengue virus in 0.22 um filters.
21 July 2016 2,242 2 View
I have a mixture of monomer and dimer. Monomer of the protein has N terminal His tag and i did not remove the his tag after purification. I provide particular condition to my monomers so that two...
21 July 2016 5,413 3 View
cell type - human fibroblasts
15 July 2016 3,370 1 View
Hi all, I design the AS-PCR assay to genotype the SNP, plasmids were used to guarantee the three cluters. However, it is strange that the hetorozygous plasmid went to wild-type area, while the...
26 June 2016 5,483 2 View