Zhenyu Shen

11 Questions 6 Answers 0 Followers

Questions related from Zhenyu Shen

use a RNA fragment to construct a standard curve for a RT-PCR?

Can I use a synthesised RNA fragment containing the amplification target to construct a standard curve for a one-step RT-PCR? Can I also use the RNA fragment as the amplification control? What...

04 April 2019 3,889 5 View

Sanger sequencing is longer than amplicon?

Sometime my Sanger sequencing is longer than the amplicon seen on the gel. Have you meet the problem? What can cause this?

02 February 2019 810 2 View

Sequence LNA PCR product?

I am doing genotyping PCR. The probe has 3 LNA nt. Can I do regular Sanger sequencing to the PCR product to verify the genotype?

11 November 2018 9,457 0 View

A short amplicon got a long sequence by Sanger sequencing?

By PCR I got a ~500 bp amplicon from a microorganism. I did gel purification and sent for Sanger sequencing with the PCR primers from both ends. Now the sequencing result from the forward primer...

06 June 2018 3,043 3 View

Primer and probe TM of one-tube real-time nested PCR?

I am designing a one-tube nested real-time PCR. Only one probe will be used and the signal will be only collected from the second stage. Any suggestions for primer and probe TM design?

05 May 2018 7,300 5 View

One primer specific?

I am designing a Taqman PCR. However, I only can find a small continue specific fragment (9bp) in the target region. How should I design primers and probe? 1. place the 9 bp in either forward...

05 May 2018 8,754 6 View

Dose SYBR Green change amplicon running speed in an agrose gel ?

As mentioned in the title, has someone compared the running speeds between SYBR Green-DNA polymerase - amplicon and conventional polymerase -amplicon in an agrose gel?

08 August 2017 4,862 1 View

QPCR amplicon did not have the band on the agrose gel

I did a QPCR, which gave me the CT around 20 and it had a sharp peak around 80 degree according to the melting curve. Then, I realized I need to extract DNA from it. So, I ran i5 ul of the 25 ul...

08 August 2017 8,523 3 View

Forward sequencing and reverse sequencing have different results

I have a ~700 PCR amplicon sequenced from both directions. However, forward sequencing and reverse sequencing have different results. The forward one has almost perfect matches with existing...

07 July 2017 6,818 4 View

How do you search primer specificity?

BLAST primer, ipress, MFEprimer ? which one do you use?

01 January 2017 2,460 0 View

How to find a genus or species specific gene for diagnosis?

For example, I know a bacterial genome. How should I search the genome with all bacterial genomes to find out a specific segment which can be used in PCR to diagnose the presence of the bacterium...

01 January 2017 3,928 4 View