Jeroen Vierstraete

8 Questions 5 Answers 0 Followers

Questions related from Jeroen Vierstraete

Cloning: BP reaction of large fragment: only a smaller fragment is found?

I perform long range-PCR of a 9kb fragment and purify the 9kb band from a gel. However after doing multiple BP reactions, i keep getting a smaller 3kb fragment that is in my plasmid, and no 9kb...

30 November 2017 8,797 3 View

Can I clean my bench with ethanol to remove RNase activity?

I am wondering if cleaning my bench with ethanol is sufficient to remove RNase activity. Or should I use products like RNase ZAP?

05 October 2017 2,161 1 View

For in vitro transcription purposes, should I use sigma or DEPC treated water to avoid RNA degradation?

I want to perform in vitro transcription of large fragments (>10kb). I am concerned about the stability of my mRNA strands so I want to avoid degradation at all costs. I am using the mMessage...

20 September 2017 7,709 6 View

Multiple bands after RNA transcription of a large gene?

I see multiple bands after performing transcription of a large gene and letting ut run over a denaturing RNA gel. Does anyone know possible causes and how to get rid of the unwanted smaller bands?

08 March 2017 4,481 3 View

Help on Gateway Cloning a large PCR fragment (10kb)?

I am currently trying to Gateway clone a large PCR fragment. I have been able to PCR it perfectly and purify it through gel extraction. However, after BP reaction and transformation I have no...

13 December 2016 5,064 3 View

Does anyone have a good protocol for obtaining large cDNA fragments (>10kb) through RT-PCR?

I want to amplify the cDNA sequence of a large gene in zebrafish (around 10kb). I am looking for any tips around RNA extraction protocol and cDNA synthesis so that my whole cDNA fragment isn't...

16 November 2016 4,855 3 View

Does the RNeasy mini kit allow for RNA extraction of large fragments (>10kb)?

Hello, I want to know if anyone has been able to get large RNA fragments from performing RNA extraction through the RNeasy mini kit. Many thanks! Jeroen

16 November 2016 2,541 5 View

Can I freeze my product after performing a restriction digest?

Is it possible for me to freeze my product after performing a restriction digest so that I can continue working tomorrow? Or is it better to place the product at 4°C?

03 December 2015 8,124 11 View