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Questions related to Molecular Biological Techniques
I prepared a 50 mM phosphate buffer in H2O and pH'ed it to 7. I lyophilized it and dissolve it in D2O? What is the pD? And what would be the pT (I would like a tritium water solution, ultimately).
03 August 2016 1,335 3 View
Is there anything I can do to reduce the amount of fuzzy and extra bands that appear behind my DGGE? Its making it difficult to differntiate between the bands I need. The standards are located at...
03 August 2016 8,067 4 View
I will be running my first luciferase assay and the protocol I am using does not specify what filters or filter settings I should use. The plate will be read on a lumometer from Biotek, Synergy...
02 August 2016 2,982 1 View
I expressed a protein in SUMO vector that express SUMO fusion protein, so, I wanna do an enzymatic essay after the protein purification without cleavage the SUMO protein. Has someone experience in...
02 August 2016 7,399 5 View
I made the siRNA and Lipofectamine2000 mix and incubated 20 minutes @ room temperature. The problem now is the cells are not healthy to start the transfer. Anybody please let me know how long i...
02 August 2016 4,600 3 View
I've been trying to clone a gene from H. influenza. Every step (PCR, digestion, ligation) seemed to work based on agarose gel images. Before I sent the DNA for sequencing, I digested the plasmids...
02 August 2016 10,052 5 View
Why C-terminal tagging of GFP did not result in the higher level of the protein expression compare to N-terminal tagging?
02 August 2016 1,602 3 View
Hi all, I am currently ligating 1.3 kb gene to pET29 vector, however, i cant get any bands after colony PCR using T7/T7t primers. I gotten a clear one band in RE digestion for clean cut vector and...
01 August 2016 9,848 4 View
Are all the bands plasmids itself?or is it degraded plasmid?
31 July 2016 7,216 11 View
Total Protein: 6His+MBP+TEV+tagret Protein: pI=6.49, MW=165.315 kDa,Cleaved Protein: target Protein: pI=8.42, MW=121.625 kDadialysis buffer: 20 mM HEPES pH 7.5, 50 mM NaCl, 10% glycerol. (similar...
29 July 2016 1,833 7 View
Does anyone know the best way to visualize ssRNA in gel electrophoresis?
29 July 2016 5,636 4 View
please also suggest articles regarding the same.
29 July 2016 1,231 1 View
Recently we have a condensate in our incubator. I'm worried how will it affect the cells, since it will definitely change the humidity levels in the chamber. Did anyone have this problem? I don't...
29 July 2016 8,593 4 View
Is cell lysis protocol same for all proteins?.
29 July 2016 1,105 3 View
I am new to PCR and digging into learning the roles of the different components in a PCR based on the FastStart HF PCR System I am using from Roche. If EDTA is a chelating agent, and we need the...
28 July 2016 784 4 View
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 1,490 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 7,795 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 5,174 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 5,090 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 9,925 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 1,869 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 2,612 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 9,148 5 View
Any suggestions are more than welcome.
26 July 2016 8,311 3 View