Leiden Muscular Dystrophy pages^©

Primers for DYSF analysis

(last modified Febuary 04, 2004)

• DNA-analysis
• RNA-analysis
  • Aoki et al. (2001)
  • Ueyama et al. (2001)

DNA-primers

Legend:
^a Reported as tcgagctcttgtcctgccAtgc by McNally but this does not match with the genomic DYSF sequence.
Exonic sequences are in upper case, intronic and gene flanking sequences in lower case and added primer tails in italics - not yet verified). Exon: exon amplified. Numbering of exons is according to Aoki et al [2001]. Length (in bp): length of PCR-product in basepairs. Forward / reverse primer: sequence of forward and reverse primer. Reference: publication describing the primer(s). Name: name of the primers. Tm (^oC): annealing temperature of the primer.

RNA-primers

• primers Aoki et al. (2001)
• primers Ueyama et al. (2001)

PCR conditions Aoki et al. (2001)

• first round RT-PCR - 3 min 94^oC (denaturation), 35 cycles of 30 sec 94^oC/ 30 sec 53^oC / 1 min 72^oC
• second round PCR - 3 min 94^oC (denaturation), 30 cycles of 30 sec 94^oC / 30 sec 56^oC / 1 min 72^oC

Legend:
First round RT-PCR / 2^nd round PCR: RT-PCR sets described by Aoki et al. [2001]. Location: position of primer sequences in relation to the DYSF-cDNA Reference Sequence. Numbering of exons is according to Aoki et al. [2001]. Length (in bp): length of amplified segment in base pairs. Forward / reverse primer: sequence of forward and reverse primer. Name: name of the primers. Reference: publication describing the primer(s).

 PCR conditions Ueyama et al. (2001)

• RT-PCR sets A-C, I and J: 2 min 94^oC (denaturation), 35 cycles of 1 min 94^oC/ 1 min 55^oC sets/ 1 min 72^oC
• RT-PCR sets D-H: 2 min 94^oC (denaturation), 35 cycles of 1 min 94^oC/ 1 min 57^oC sets / 1 min 72^oC

Legend:
RT-PCR set: RT-PCR sets described by Ueyama et al. [2001]. Location: position of primer sequences in relation to the DYSF-cDNA Reference Sequence (NOTE: location nor sequence has been verified yet). Numbering of exons is according to Aoki et al. [2001]. Length (in bp): length of amplified segment in base pairs. Forward / reverse primer: sequence of forward and reverse primer. 

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