PCR, Cloning, Restriction Digestion, Ligation, Transformation, Plasmid et al | Contact experts in Molecular Biology to get answers
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Questions related to Molecular Biology
We have tried to make a lyophilized PCR mixture. However, when the elegant cakes are rehydrated with target samples, small bubbles are cosiderably generated coincidentally. How can we reduce the...
27 July 2017 9,933 3 View
Hi everyone I start to get the work about genomic DNA PCR, I need some suggestions such as PCR program setting, Taq DNA polymerase, or sample preparing, or...... The size of amplicons were about...
26 July 2017 1,421 3 View
I am trying to do in vitro digestion of DNA with Cas9 Nuclease, S. pyogenes (M0396 NEB). When I run the DNA on a gel, the DNA is smearing instead of showing up as clear bands. I examined the gel...
26 July 2017 8,560 6 View
I have some primers that were initially designed for a TaqMan qPCR but in their simple version and I want to use them for an end point PCR. What kind of program should I use? Thank you, Alkmini
26 July 2017 2,198 0 View
If you require further details before answering please let me know, thanks
26 July 2017 8,494 2 View
Hi everyone! I am planning to co-express two proteins in insect cells using the pFastBac Dual expression vector. I have a question regarding the multiple cloning site upstream of the p10 promoter....
26 July 2017 219 1 View
I have done RNA isolation from a kidney cell line ( RIN 9.3), followed by reverse transcription using 1ug of RNA using iscript kit. But after qPCR, I found the positive control Ct value is 29...
26 July 2017 999 6 View
Does anyone have a protocol recommendation for digesting a collagen matrix with MMP-1?
25 July 2017 8,145 2 View
I am using the Smarter kit from Clontech to know 5' and 3' ends of some transcripts. I have done the controls PCR (TFR primers) but I only get band with the 3' end reaction. Does anybody know what...
25 July 2017 6,709 1 View
I have to clone a plasmid in which the gene of interest will be expressed by a PGK1 promoter. Can I use a human PGK1 promoter when I want to express my protein in mouse cells? This would greatly...
25 July 2017 1,376 3 View
use for in vivo experiment.
24 July 2017 3,947 2 View
I am working on one gene having 19 exons with 4 transcripts. A deletion mutation (one bace pair deletion) is present in all the four transcripts of this gene. It leads to frameshift and make a...
24 July 2017 8,745 8 View
The principle of Ampure XP beads
24 July 2017 8,761 4 View
Hello everybody, I´m a bachelor´s student I nee dto amplify a 5 kb fragment from a plasmid. Therefore, I perform PCR, and then gel electrophoresis. Subsequently, I extract the 5 kb band and try to...
24 July 2017 3,452 4 View
I have extracted a sample using do-decane (5ml) and need to concentrate the sample for GC analysis.
24 July 2017 480 2 View
Hello! I am microinjecting Tol2 and UAS:mScarlet into zebrafish, and I was looking up what the optimal concentration of Tol2 and plasmid I should be injecting. There were a couple protocols that...
24 July 2017 8,976 4 View
I want to do SYBR real time PCR for relative quantification of TGFB1. Gel electrophoresis of real time PCR product showed two bands in around about 80 bp (79 oC melting peak) and 180 bp (84 oC...
23 July 2017 2,359 1 View
Dear All, I have encountered some problem in Gibson Cloning. The primer was ordered from Sigma with melting temperature at 65'C and the PCR for both linearizing the both vector (3500kb) and...
22 July 2017 822 3 View
I have 1kb desired amplified product but after column or Na-acetate purification i am getting ~700bp band along with this desired band now with less intensity. Cannot understand what is happening....
22 July 2017 7,114 4 View
I am looking to digest a bovine collagen matrix and MMP-1 continues to be recommended over bacterial collagenase. Is there a specific reason for this?
21 July 2017 5,669 1 View
i regulaarly grow in 28 degers in wild type but i mutate alanine to cystine if it grow in 28 it goes to pellet 0.5mM IPTG 5h after that i cheaked 18 degres 0.1mM IPTG 12h the yield little bit...
21 July 2017 5,181 2 View
If small RNAs are originated from one source with different lengths (20-24nt), how can I distinguish them? Is Stem-loop PCR method useful to distinguish them?? Any suggestion is appreciated. Thank you
21 July 2017 4,436 9 View
analysis of TDT
21 July 2017 7,356 1 View
We are planning to buy shRNA for our experiments. can anyone suggest which form is good like whether one should buy bacterial stocks, plasmid or lentiviral particles? Is it good to buy bacterial...
21 July 2017 4,481 3 View